Hybrid toxin

ABSTRACT

The present invention provides, inter allia, a B.t. hybrid toxin fragment comprising at its C-terminal domain III of a first Cry protein or a part of the domain or a protein substantially similar to the domain, with the proviso that the N-terminal region of the fragment is the N-terminal region of a second Cry protein or a part of the region or a protein substantially similar to the region.

The present invention relates to hybrid toxin figments, and toxinscomprising them, derived from Bacillus thuringiensis insecticidalcrystal proteins.

Bacillus thuringiensis (hereinafter B.t.) is capable of producingproteins that accumulate intracellularly as crystals. These crystalproteins are toxic to a number of insect larvae. Based on sequencehomology and insecticidal specificity, crystal proteins have beencategorized into different classes. Best studied are the CryI class ofproteins which are produced as 140 kDa pro-toxins and are active towardslepidopterans.

To some extent the mode of action of crystal proteins has beenelucidated. After oral uptake the crystals dissolve in the alkalineenvironment of the larval midgut. The solubilized proteins aresubsequently processed by midgut proteinases to a proteinase-resistanttoxic fragment of about 65 kDa which binds to receptors on epithelialcells of the insect midgut and penetrates the cell membrane. Thiseventually leads to bursting of the cells and death of the larvae.

The activity spectrum of a particular crystal protein is to a largeextent determined by the occurrence of receptors on the midgutepithelial cells of susceptible insects. The said spectrum isco-determined by the efficiency of solubilization of the crystal proteinand its proteolytic activation in vivo.

The importance of the binding of the crystal protein to midgutepithelial receptors is further demonstrated where insects havedeveloped resistance to one of the crystal proteins in that the bindingof crystal proteins to midgut epithelial cells in resistant insects issignificantly reduced.

Toxic fragments of crystal proteins are thought to be composed of threedistinct structural domains. Domain I, the most N-terminal domain,consists of 7 α-helices. Domain II comprises 3 β-sheets and domain III(the most C-terminal) folds into a β-sandwich. If projected on CryIsequences, domain I runs from about amino acid residue 28 to 260; domainII from about 260 to 460 and domain III from about 460 to 600.

The present invention concerns hybrid crystal proteins particularly,though not exclusively, involving CryIC and CryIE or CryIA. Thenucleotide sequence of the CryIC gene from B.t. sub.sp. entomocidus 60.5is given in SEQ ID. No. 1, and the corresponding amino acid sequence ofthe protein encoded by the said nucleotide sequence is given in SEQ IDNo. 2. The nucleotide sequence of the CryIE gene from B.t. sub. sp.kenyae 4FI is given in SEQ ID. No. 3, and the corresponding amino acidsequence of the protein encoded by the said nucleotide sequence is givenin SEQ ID No. 4. These proteins are toxic to lepidopterans, but withinthis order of insects, each protein has different specificity. CryIC isparticularly active against S. exigua and M. brassicae.

According to the present invention there is provided a B.t. hybrid toxinfragment comprising at its C-terminal domain III of a first Cry proteinor a part of the domain or a protein substantially similar to thedomain, with the proviso that the N-terminal region of the fragment isthe N-terminal region of a second Cry protein or a part of the region ora protein substantially similar to the region. A preferred fragment isone which does not bind to the CryIC binding site in an insect gut whenit comprises at its C-terminal domain III of CryIC or a part of thedomain or a protein substantially similar to the domain; or one whichdoes not bind to a CryIA binding site when it comprises at itsC-terminal domain III of CryIA or a part of the domain or a proteinsubstantially similar to the domain.

By substantially similar is meant pure proteins having an amino acidsequence which is at least 75% similar to the sequence of the proteinsaccording to the invention. It is preferred that the degree ofsimilarity is at least 85%, more preferred that the degree of similarityis at least 90% and still more preferred that the degree of similarityis at least 95%.

In the context of the present invention, two amino acid sequences withat least 75%, 85%, 90% or 95% similarity to each other have at least75%, 85%, 90%, or 95% identical or conservatively replaced amino acidresidues in a like position when aligned optimally allowing for up to 6gaps with the proviso that in respect of the gaps a total not more than15 amino acid residues are affected. For the purpose of the presentinvention conservative replacements may be made between amino acidswithin the following groups:

(i) Serine, and Threonine;

(ii) Glutamic acid and Aspartic acid;

(iii) Arginine and Lysine;

(iv) Asparagine and Glutamine;

(v) Isoleueine, Leucine, Valine and Methionine;

(vi) Phenylalanine, Tyrosine and Tryptophan

(vii) Alanine and Glycine

with the proviso that in SEQ ID No. 6 Ser and Tyr are conservativereplacements at position 620, and Ala and Glu are conservativereplacements at position 618; and that in SEQ ID No. 8 Ser and Tyr areconservative replacements at position 627, and Ala and Glu areconservative replacements at position 625.

By "part" of a protein is meant a peptide comprised by the said proteinand having at least 80% of the consecutive sequence thereof.

By "binding site" is meant a site on a molecule wherein the bindingbetween site and toxin is reversible such that the Ka between site andtoxin is in the order of at least 10⁴ dm³ mole⁻¹.

The toxin fragment may comprise at its N-terminal the N-terminal regionof any insecticidal proteins from B.t. being commonly known as "Cry" or"Cyt", including: CryIA(a), CryIA(b) CryIA(c), CryIB, CryIC, CryID,CryIE, CryIF, CryIG, CryIH, CryIIA, CryIIB, CryIIC, CryIIIA, CryIIIB,CryIIIB(b), CryIVA, CryIVB, CryIVC, CryIVD, CYTA, CryX1(IIIC),CryX2(IIID), CryX3, CryV, and CryX4; or a part of the region, or aprotein substantially similar to the region, and that the C-terminal ofthe fragment is domain III of CryIC or a part of the domain or a proteinsubstantially similar to the domain.

Thus the fragment may comprise domain II of CryIE, CryIB, CryID orCryIA, or a part of the domain or a protein substantially similar to thedomain, and domain III of CryIC or a part of the said domain III or aprotein substantially similar to the said domain III. It is particularlypreferred that the fragment comprises domains I and II of CryIE, CryIB,CryID or CryIA or a part thereof or a protein substantially similar tothe said domains, and domain III of CryIC or a part thereof or a proteinsubstantially similar to the said domain III.

It is most preferred that the toxin fragment comprises a region at itsC-terminus comprising the sequence from amino acid position 454 toposition 602 of CryIC, or a sequence substantially similar to the saidsequence. The fragment may comprise a region at its C-terminuscomprising the sequence from amino acid position 478 to 602 of Cry IC,or a sequence substantially similar to the said sequence, with theproviso that if the sequence comprising amino acids 478 to 602 of CryICis fused directly to the C-terminus of domain II of CryIA, CryIB, CryIDor CryIE then the folding of the fusion product is satisfactory to yieldan insecticidal component of the fragment. The skilled man willrecognize that it may be necessary to add a peptide region to theC-terminus of domain II which spaces the C-terminal region of CryICapart thus enabling it to fold in such a way as to exhibit insecticidalactivity.

It is most particularly preferred that the toxin fragment according tothe invention comprises either:

i) an amino acid sequence from about amino acid 1 to about amino acid620 in SEQ ID No. 6, or an amino acid sequence from about amino acid 1to about amino acid 620 in SEQ ID No. 6, wherein with respect to thesaid sequence, at least one of the following alterations is present:

Ile at position 609 is replaced with Leu;

Ala at position 618 is replaced with Glu;

Ser at position 620 is replaced with Tyr, or

ii) an amino acid sequence from about amino acid 1 to about amino acid627 in SEQ ID No. 8, or an amino acid sequence from about amino acid 1to about amino acid 627 in SEQ ID No. 8, wherein with respect to thesaid sequence, at least one of the following alterations is present:

Ile at position 616 is replaced with Leu;

Ala at position 625 is replaced with Glu;

Ser at position 627 is replaced with Tyr.

Whatever amino acid alterations are permitted, however, one or more ofthe following residues--indicated sequence-wise with respect to theCryIC sequence--is invariable: Phe (501); Val (478) Trp (479) and Thr(486).

The invention also includes a hybrid toxin comprising the abovedisclosed fragment or a toxin at least 85% similar to such a hybridtoxin which has substantially similar insecticidal activity, or receptorbinding properties.

The invention still further includes pure proteins which are at least90% identical to the toxin fragments or hybrid toxins according to theinvention.

The invention still further includes recombinant DNA comprising asequence encoding a protein having an amino acid sequence of the abovedisclosed toxins or fragments thereof. The invention still furtherincludes recombinant DNA comprising the sequence from about nucleotide 1to about nucleotide 1860 given in SEQ ID. No. 5 or DNA similar theretoencoding a substantially similar protein, or recombinant DNA comprisingthe sequence from about nucleotide 1 to about nucleotide 1881 in SEQ IDNo. 7 or DNA similar thereto encoding a substantially similar protein.

By similar DNA is meant a test sequence which is capable of hybridizingto the inventive recombinant sequence. When the test and inventivesequences are double stranded the nucleic acid constituting the testsequence preferably has a TM within 20° C. of that of the inventivesequence. In the case that the test and inventive sequences are mixedtogether and denatured simultaneously, the TM values of the sequencesare preferably within 10° C. of each other. More preferably thehybridization is performed under stringent conditions, with either thetest or inventive DNA preferably being supported. Thus either adenatured test or inventive sequence is preferably first bound to asupport and hybridization is effected for a specified period of time ata temperature of between 50° and 70° C. in double strength citratebuffered saline containing 0.1%SDS followed by rinsing of the support atthe same temperature but with a buffer having a reduced SCconcentration. Depending upon the degree of stringency required, andthus the degree of similarity of the sequences, such reducedconcentration buffers are typically single strength SC containing0.1%SDS, half strength SC containing 0.1%SDS and one tenth strength SCcontaining 0.1%SDS. Sequences having the greatest degree of similarityare those the hybridization of which is least affected by washing inbuffers of reduced concentration. It is most preferred that the test andinventive sequences are so similar that the hybridization between themis substantially unaffected by washing or incubation in one tenthstrength sodium citrate buffer containing 0.1%SDS.

The recombinant DNA may further encode a protein having herbicideresistance, plant growth-promoting, anti-fungal, anti bacterial,anti-viral and/or anti-nematode properties. In the case that the DNA isto be introduced into a heterologous organism it may be modified toremove known mRNA instability motifs (such as AT rich regions) andpolyadenylation signals, and/or codons which are preferred by theorganism into which the recombinant DNA is to be inserted may be used sothat expression of the thus modified DNA in the said organism yieldssubstantially similar protein to that obtained by expression of theunmodified recombinant DNA in the organism in which the proteincomponents of the hybrid toxin or toxin fragments are endogenous.

The invention still further includes a DNA sequence which iscomplementary to one which hybridizes under stringent conditions withthe recombinant DNA according to the invention.

Also included in the present invention are: a vector containing such arecombinant (or complementary thereto) DNA sequence; a plant ormicro-organism which includes, and enables expression of such DNA;plants transformed with such DNA; the progeny of such plants whichcontain the DNA stably incorporated and hereditable in a Mendelianmanner, and/or the seeds of such plants and such progeny.

The invention still further includes protein derived from expression ofthe said DNA, and insecticidal protein produced by expression of therecombinant DNA within plants transformed therewith.

The invention still further includes an insecticidal compositioncontaining one or more of the toxin fragments or toxins comprising themaccording to the invention; a process for combatting insects whichcomprises exposing them to such fragments or toxins or compositions, andan extraction process for obtaining insecticidal proteins from organicmaterial containing them comprising submitting the material tomaceration and solvent extraction.

The invention will be further apparent from the following description,which describes the production of B.t. hybrid toxin fragments accordingto the invention, taken in conjunction with the associated drawings andsequence listings.

FIG. 1 shows the generation of hybrid crystal protein genes via in vivorecombination. Tandem plasmids (pBD560 and pBD 650) carrying twotruncated crystal protein genes in direct repeat orientation areconstructed. The 5' located gene (open bar) lacks the protoxin encodingregion (solid bar) and of the 3' located gene (dashed bar) part of thedomain I encoding region is deleted. In vivo recombination betweenhomologous regions (domain II and III) occurs in recA+ strain JM101.Selection against non-recombinants by digestion with NotI and BamHI andsubsequent transformation results in sets of plasmids encoding hybridcrystal proteins.

FIG. 2 shows the alignment of amino acid residues 420 to 630 of CryIEand CryIC. The border between domain II and III is indicated. Only aminoacid residues of CryIC which differ from CryIE are depicted, identicalresidues are indicated by a dot. The positions of cross-over (G27, H13,H7, H8 H17 and H21) in the CryIE-CryIC hybrid toxin fragments accordingto the invention are indicated on the Figure.

FIG. 3 shows the alignment of amino acid residues 420 to 630 of CryIEand CryIC. The border between domain II and III is indicated. Only aminoacid residues of CryIC which differ from CryIE are depicted, identicalresidues are indicated by a dot. The positions of cross-over (F59, F71,F26, and E7) in the CryIC-CryIE hybrid toxin fragments are indicated onthe Figure.

FIGS. 4a and 4b show the results of some heterologous competitionexperiments. Biotinylated CryIC (FIG. 4a) and G27 (FIG. 4b) areincubated with S. exigua BBMV vesicles in the absence (lanes a) orpresence of an excess of unlabelled protein as indicated. After theincubation, the vesicles are washed, loaded on a SDS-polyacrylamide geland blotted to a nitrocellulose membrane. Biotinylated crystal proteins,re-isolated with the vesicles, are visualized usingstreptavidin-peroxidase conjugate.

FIG. 5 shows the plasmid map of pSB456 which encodes the G27 hybridtoxin fragment and is used to transform the crystal toxin minus strainB.t. 51.

SEQ ID. No. 1 shows the nucleotide sequence of the CryIC gene from B.t.sub.sp. entomocidus 60.5.

SEQ ID No. 2 shows the amino acid sequence of the protein encoded by theCryIC gene shown in SEQ ID No. 1.

SEQ ID No. 3 shows the nucleotide sequence of the CryIE gene from B.t.sub.sp. kenyae 4FI.

SEQ ID No. 4 shows the amino acid sequence of the protein encoded by theCryIE gene shown in SEQ ID No. 3.

SEQ ID No. 5 shows the nucleotide sequence encoding a preferredCryIE/CryIC B.t. hybrid toxin fragment according to the invention.

SEQ ID No. 6 shows the amino acid sequence of the protein encoded by thenucleotide sequence shown in SEQ ID No. 5.

SEQ ID No. 7 shows the nucleotide sequence of a CryIA/CryIC hybrid toxinfragment according to the invention.

SEQ ID No. 8 shows the amino acid sequence of the protein encoded by thenucleotide sequence depicted in SEQ ID No.7.

Production of plasmids encoding Hybrid toxin fragments

In the production of plasmids carrying the CryIC or CryIE genes,Escherichia coli XLI-blue (Stratagene Inc.) is used as plasmid hostexcept in cases were JM101 is used as recA+ background. A vector for theexpression of crystal proteins in E. coli is derived from pKK233-2(Pharmacia LKB Biotechnology). The size of pKK233-2 is reduced bydeleting an EcoRI-PvuII fragment carrying the gene encoding tetracyclineresistance. Subsequently a 6 bp XhoI linker is ligated into the HindIIIsite resulting in pBD10. Plasmid BK+ is created by insertion of a BglIIlinker in the SacI site of Bluescript SK+ (Stratagene Inc.). Thepolylinker of BK+ from BglII to XhoI is introduced between the NcoI-XhoIsite in pBD10. The resulting expression vector pBD11 contains the highlyexpressed trc promoter, the lacZ ribosome binding site and ATGinitiation codon. The initiation codon overlaps with a NcoI site and isfollowed by the polylinker to facilitate insertions into the vector.Transcription is terminated by the rrnB transcription terminator.

The cloning of the cryIC and cryIE genes from B.t. subsp. entomocidus60.5 and kenya 4F1 respectively, is as described previously (Honee etal., 1990 (Appl. Environ. Microbiol. 56, pp 823-825); Visser et al.,1990 (J. Bacteriol. 172, pp 6783-6788)). For cloning purposes, an NcoIsite overlapping with the start codon of cryIC is created by in vitromutagenesis. A BglII site is created directly downstream of thetranslation termination codon of cryIC by site directed mutagenesis,resulting in the sequence ATAAGATCTGTT (stop-codon underlined). TheNcoI-BglII fragment containing the cryIC coding region is ligated intopBD11, resulting in CryIC expression plasmid pBD150. pBD155 is aderivative of pBD150, in which the polylinker sequences 3' of cryIC aredeleted.

A DraI fragment from pEM14 (Visser et al., 1990) containing the completecryIE gene is cloned in the EcoRV site of SK+, resulting in plasmidpEM15. Subsequently, an NcoI site is introduced by site directedmutagenesis at the startcodon of the gene, and cryIE is transferred asan NcoI-XhoI fragment to pBD11 resulting in CryIE expression plasmidpBD160.

Plasmids carrying only toxic fragment encoding regions of the cryI genesare constructed. BglII linkers are ligated to XmnI sites present at bpposition 1835 of cryIC, and to the HgiAI site at position 1839 of cryIE.Subsequently, NcoI-BglII fragments containing the cryIC (1835 bp) andcryIE (1839 bp) toxic fragment encoding regions are ligated into pBD11resulting in pBD151 and pBD161 respectively as described below.

Tandem plasmids used for the generation of cryIC-cryIE hybrid genes areconstructed as follows. BamHI linkers are ligated to pBD160 digestedwith HpaI. This DNA is incubated with BamHI and XhoI and the truncatedcryIE gene running from bp 704 is ligated into pBD151 resulting inpBD560. To construct a tandem plasmid for the generation of cryIE-cryIChybrids, pBD155 is digested with NsiI and XhoI. The fragment carryingthe truncated cryIC gene, running from bp 266, is ligated into PstI/XhoIdigested pBD161, resulting in plasmid pBD650. Due to polylinkersequences, unique NotI and BamH1 restriction sites are present betweenthe truncated cryI genes present in the tandem plasmids pBD560 andpBD650.

DNA manipulations and construction of hybrid toxins.

All recombinant DNA techniques are as described by Sambrook et al. 1989(in "Molecular Cloning", A Laboratory Manual: Cold Spring Harbour Press,Cold Spring Harbour), DNA sequencing is performed by thedideoxytriphosphate method with fluorescent dyes attached to thedideoxynucleotides. Analysis is automated by using an Applied Biosystems370A nucleotide sequence analyzer.

The homology present between cryI genes permits intramolecularrecombination in vivo. Two tandem plasmids are created by, each carryingtwo truncated crystal protein genes overlapping only in domains II andIII. Therefore, recombination occurs only in regions encoding domains IIand III. In frame recombinations, which can be selected for byrestriction enzyme digestion, generate plasmids which express full size140 kDa hybrid protoxins. To generate in vivo recombinants, a tandemplasmid (either pBD560 or pBD650; FIG. 2) is transferred to JM101. 5 μgof DNA is isolated from independently generated recombinants and isdigested with NotI and BamHI cutting between the two truncated cryIgenes to select against non-recombinants and the DNA is transformed toE. coli XL1-blue. 5 single colonies are grown and protein patterns andplasmid content are analysed. CryIC-CryIE and CryIE-CryIC hybrid toxinsare generated using the tandem plasmids pBD560 and pBD650 respectivelywhich are allowed to recombine in a recA+ background, DNA is isolated,digested and transferred to recA- strain as described above.

100 colonies of 20 independent experiments are analyzed on SDS-PAGE. 85%of these clones produce a 140 kDa protein indicating in framerecombinations between cryIC and cryIE, and cryIE and cryICrespectively. In E. coli, CryI proteins are produced as crystals whichcan be solubilized in vitro at high pH. Approximately 15% of hybridtoxins produced as above are solubilized at high pH. The recombinantsproducing soluble hybrid toxins are first classified using restrictionenzymes, subsequently for each class the cross-over point of selectedhybrids is determined by DNA sequence analysis. All cross-overs whichresulted in soluble hybrid toxins occur in or very close to domain III.

Protein purification and analysis.

Crystal proteins are isolated essentially as described by Convents et al(J. Biol. Chem. 265, pp 1369-1375; Eur. J. Biochem, 195, pp 631-635).Briefly, recombinant E. coli are grown at 30° C. in 250 ml TB medium toan OD₆₀₀ of 10-15. Crystals isolated from the E. coli lysate aresolubilized during incubation for 2 h in 20 mM Na₂ CO₃, 10 mMdithiothreitol, 100 mM NaCl, pH10 at 37° C. The pH of the solution islowered to 8 with Tris-HCl and incubated with trypsin. The toxinsolution is dialysed against 20 mM Tris-HCl, 100 mM NaCl pH9.Subsequently the toxic fragment is purified on a Mono Q 5/5 columnconnected to a fast-protein liquid chromatography (FPLC) system(Pharmacia LKB Biotechnology). Proteins are separated by 7.5% sodiumdodecyl sulfate-polyacrylamide gel electrophoreses.

Biochemical analysis and isolation of 65 kDa toxic fragments.

Isolated crystals of purified CryIC, CryIE and the hybrid proteins aresolubilized at high pH and incubated with trypsin. Like CryIC and CryIE,all soluble hybrid toxins with cross-overs in domain III are convertedto stable 65 kDa fragments. The 65 kDa fragments can be purified usinganion exchange chromatography under similar conditions as the parentalproteins. Hybrids F59 and F71 which have cross-overs in domain II, arecompletely degraded by trypsin. Apparently, although these hybrids donot precipitate as insoluble aggregates, trypsin cleavage sites buriedin the parental proteins may become exposed to trypsin. Because of thisphenomenon, no 65 kDa fragments are isolated from F59 and F71.

Table 1 shows the constitution of 5 CryIE-CryIC hybrid toxins: (G27; H8;H17; H13; H7 and H21) and 4 CryIC-CryIE hybrid toxins (F59; F71; F26;E7) with reference to the CryIC and CryIEE proteins from which they arederived. The amino acid sequences of the CryIE-CryIC toxins comprisingthe toxic fragments of the present invention run to amino acid 1189 ofthe CryIC parent protein. The amino acid sequences of the CryIC-CryIEhybrid toxins run to amino acid 1171 of the CryIE parent protein. Table1 also shows the relative insecticidal effectiveness of these varioushybrid toxins with respect to the CryIC and CryIE proteins.

                  TABLE 1                                                         ______________________________________                                                                                M.                                    TOX. a.a IE      a.a. IC  M. sexta                                                                             S. exigua                                                                            brassicae.                            ______________________________________                                        IC   0           28-627   ++     ++     ++                                    IE   29-612      0        ++     -      -                                     G27  1-474       478-627  ++     ++(+)  +(+)                                  H8   1-497       501-627  ++     -      -                                     H17  1-529       533-627  ++     -      -                                     H7   1-577       588-627  -      -      -                                     H21  1-605       621-627                                                      F59  421-612     1-423    -      -      -                                     F71  428-612     1-430    -      -      -                                     F26  455-612 (1171)                                                                            1-458    ++     -      -                                     E7   588-612 (1171)                                                                            1-602    ++     ++     ++                                    ______________________________________                                         Table 1. Constitution and toxicity of hybrid toxins with respect to the       parent proteins. Most bioassays were performed with purified toxin            fragments. In case of cryIC these run from about aa 28 to about aa 627,       and in case of cryIE till 612. The length of complete protoxins is            indicated between brackets.                                              

Insect toxicity assays and insecticidal activity of cryIC/cryIE hybridgene products.

Bacterial cultures are concentrated to OD₆₀₀ 6.0 and 100 μl are spottedon 2 cm² of artificial diet in a 24-well tissue culture plate.Alternatively, diluted samples of purified toxins are applied to thediet. Second instar larvae of either S. exigua, M. brassicae or M. sextaare fed on this diet (16 per sample dilution) for 5 days, after whichthe larval weight is scored. The relative growth (EC50, theconcentration giving 50% growth reduction) is determined by calculatingthe ratio between the mean weight of larvae grown on diet supplementedwith toxin and the mean weight of control larvae grown on a diet withouttoxin. M. sexta egg layers are supplied by Carolina Biological SupplyCompany, North Carolina, US.

The toxic fragments encoded by the hybrid gene products are tested foractivity towards three different insect species as described above. M.sexta is susceptible to both CryIC and CryIE. As may be anticipated fromtheir sensitivity to trypsin, hybrids F59 and F71 are not active againstthis insect (Table 1). Although H7 is converted by trypsin to stable 65kDa proteins, it is not toxic to M. sexta. All of the other hybridsgiven in Table 1 are toxic and are apparently in the native,biologically active conformation.

The 65 kDa fragment of CryIC is highly toxic towards S.exigua andM.brassicae whereas CryIE is not. G27 (Table 1; FIG. 2), a CryIE-CryIChybrid with a cross-over at the junction of domain II and III is activetowards both insects. This demonstrates that domain III of CryIC confersfull activity towards S.exigua and M.brassicae. Hybrid H8, which differsin only three amino acid residues (see FIG. 3) from G27, although activeagainst M.sexta, is not active against S.exigua and M.brassicae.

F26 (Table 1, FIG. 3), the reciprocal hybrid of G27, in which domain IIIof CryIC has been exchanged by domain III of CryIE, is not activeagainst S.exigua or M.brassicae. Apparently, although the protein istoxic to M.sexta, the CryIC sequences running from amino acid 28-462 arenot sufficient to kill S.exigua and M.brassicae. Only when CryICsequences up till amino acid residue 602 are present in the hybrid (E7),is insecticidal activity against these insects restored.

The present disclosure indicates that amino acid residues from 478-602of CryIC can confer high insecticidal activity to CryIE against S.exiguaand M.brassicae.

Biotinylation of crystal proteins and binding assays.

Biotinylation is performed using biotin-N-hydroxysuccinimide esteressentially as described by the manufacturer (Amersham). 1 mg of crystalprotein is incubated with 40 μl biotinylation reagent in 50 mM NaHCO3,150 mM NaCl pH8, for one hour at 20° C. The solution is loaded on aSephadex 25 column equilibrated with the same buffer containing 0.1% BSAto remove unbound biotin and samples of the fractions are spotted on anitrocellulose membrane. Fractions containing biotinylated crystalproteins are visualized using streptavidine-peroxidase conjugate(Amersham) which catalyzes the oxidation of luminol, resulting inchemiluminescence (ECL, Amersham) and pooled.

Brush border membrane vesicles are isolated as described byWolfersberger et al. (1987) (Comp. Biochem. Physiol. 86a, pp 301-308)except that the vesicles are washed once more with isolation buffercontaining 0.1% Tween 20. Binding of biotinylated crystal proteins tobrush border membrane vesicles (100 μg/ml) is performed in 100 μl of PBScontaining 1% BSA, 0.1% Tween-20 (pH 7.6). Vesicles (20 ug vessicleprotein) are incubated with 10 ng biotinylated crystal proteins in thepresence or absence of 1000 fold excess of unlabelled crystal proteinsfor 1 hour at 20° C. Subsequently, the vesicles are re-isolated bycentrifugation for 10 minutes at 14,000 g in an Eppendorf centrifuge,washed twice with binding buffer, re-suspended in sample buffer,denatured by heating and loaded on 7.5% polyacrylamide gels. Afterelectrophoresis, proteins are blotted to nitrocellulose membranes andbiotinylated crystal proteins which are re-isolated with the vesiclesare visualized by incubation of the nitrocellulose withstreptavidin-peroxidase conjugate (Amersham) which catalyzes theoxidation of luminol, resulting in chemiluminescence (ECL, Amersham).

Since binding to epithelial gut cells is a key step in the mode ofaction of crystal proteins, the binding of crystal proteins to S. exiguabrush border membrane vesicles is investigated in heterologouscompetition experiments. Competition experiments demonstrate that thebinding of labeled CryIC (FIG. 4A, lane a) and labelled F26 (not shown)can be outcompeted by an excess of both unlabelled CryIC (lane b) or F26(lane e) but not with an excess of G27 (lane c) or CryIE (lane d).Furthermore, binding of labelled G27 (FIG. 4B, land a) and labelledCryIE (not shown) can be outcompeted by an excess of G27 (lane b) orCryIE (lane d), but not with an excess of CryIC (lane a) or F26 (lanee). From these results it is concluded that G27 and CryIE recognize thesame binding sites on S. exigua midgut membranes and that these sitesdiffer from those which are recognized by CryIC and F26. The toxicityand binding assays combined demonstrate that G27 is as toxic as CryICbut that it binds a receptor different therefrom. As insects can developresistance against a crystal protein by changing receptor bindingcharacteristics G27 may be used in resistance management programs as analternative to CryIC.

Expression of cryIE/cryIC hybrid toxin genes in heterologous systems.

The G27 cryIE/cryIC hybrid toxin gene is expressed in E. coli and thegene product exhibits at least the same insecticidal activity (at leastagainst Spodoptera) as CryIC. Moreover, the said product exhibits anincreased such activity when expressed in a Bacillus thuringiensisstrain (see below). The gene encoding the G27 hybrid toxin is introducedinto a suitable shuttle vector system, which is then introduced into anappropriate B.t. host. Such transformed cells are then cultured and theresulting toxin from both whole cultures and purified crystals isassayed for insecticidal activity.

Construction of a G27-containing shuttle vector, transformation of Bt51and purification of toxin protein therefrom.

The gene encoding hybrid G27 (3.4 kbb) is cleaved from a pKK233 E. coilexpression plasmid using Ncol and Xhol. The Xhol site is filled in usingthe Klenow fragment of E. coil DNA Polymerase I. The resulting fragmentis ligated to NcoI/Smal digested pSB635 (pBluescriptKS+, P_(crylC), andthe CryIA(c) transcription terminator). The resulting plasmid, pSB453 isdigested with Apal and Notl yielding a 4.2 kbp fragment carrying thepromoter, the hybrid G27 orf and the terminator. This fragment isligated to Apal/Notl digested pSB634 (shuttle vector containing pBC16.1and pBluescriptKS+), yielding pSB456 (see FIG. 5). Plasmid DNA, isolatedfrom E. coil DH10B, is used to transform the crystal toxin minus B.t.strain, Bt51. Positive isolates are tetracycline resistant, show thepresence of pSB456, and contain large inclusions corresponding to a 135kDa protein (as determined by SDS-PAGE. G27 hybrid toxin samples areprepared from cultures of transformed B.t.51 grown through sporulationat 30° C. in CYS-Tc¹⁰ media. Insecticidal bioassays (Table 2) areperformed on both full whole cultures and on washed crystal proteinpreparations. Controls include Bt51 (pSB440), containing the CryICtoxin, and Bt51 (pSB636), containing CryIE. Toxin concentrations areestimated by SDS-PAGE.

                  TABLE 2                                                         ______________________________________                                        Bioassay of the hybrid toxin G27 in comparison with CryIC and CryIE.          The number of samples is given in parenthesis. The hybrid toxin G27           is about 50% more effective than either of CryIE or CryIC in respect          of toxicity at least to Spodoptera sp.                                               LC.sub.50                                                              Toxin    Whole Culture (ppt)                                                                          Washed Crystal Protein (ppm)                          ______________________________________                                        CryIC    56(2)   36(2)  40(4) 7.8(2)   8.1(4)                                 CryIE    79(1)   78(1)  33(4) 11.1(6)  7.4(4)                                 G27      29(2)   21(2)  25(4) 4.7(4)   6.0(4)                                 Ratio (IC/G27)                                                                         1.93    1.71   1.60  1.66     1.35                                   ______________________________________                                    

Although the present invention has been particularly described withreference to the production of the G27 hybrid toxin, the skilled manwill appreciate that many other hybrid toxins having improvedinsecticidal characteristics may be produced according to the presentdisclosure. SEQ ID Nos. 7 and 8, for example, depict further hybridtoxins according to the invention which have improved insecticidalactivity. Hybrid toxins may be produced comprising domain III of CryICand an N-terminal of any other Cry protein. In terms of bioassays, thehybrid toxin carrying transformants may be grown in SOP media in orderto expedite the assay procedures and reduce the volumes of materialrequired. Moreover, the gene encoding G27 and/or other hybrid toxinsaccording to the invention may be transferred into toxin-encodingstrains of B.t. and/or integrated into the chromosome of selectedstrains of B.t. or indeed introduced into plant genomes to provide forinsecticidal activity in situ within the plant per se. In this regard,it is particularly preferred that the recombinant DNA encoding thetoxins is modified in that codons which are preferred by the plant intowhich the recombinant DNA is to be inserted are used so that expressionof the thus modified DNA in the said plant yields substantially similarprotein to that obtained by expression of the unmodified recombinant DNAin the organism in which the protein components of the hybrid toxin ortoxin fragments are endogenous.

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 8                                                  (2) INFORMATION FOR SEQ ID NO: 1:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 3567 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: unknown                                                         (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iii) ANTI-SENSE: NO                                                          (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Bacillus thuringiensis                                          (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..3567                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:                                      ATGGAGGAAAATAATCAAAATCAATGCATACCTTACAATTGTTTAAGT48                            MetGluGluAsnAsnGlnAsnGlnCysIleProTyrAsnCysLeuSer                              151015                                                                        AATCCTGAAGAAGTACTTTTGGATGGAGAACGGATATCAACTGGTAAT96                            AsnProGluGluValLeuLeuAspGlyGluArgIleSerThrGlyAsn                              202530                                                                        TCATCAATTGATATTTCTCTGTCACTTGTTCAGTTTCTGGTATCTAAC144                           SerSerIleAspIleSerLeuSerLeuValGlnPheLeuValSerAsn                              354045                                                                        TTTGTACCAGGGGGAGGATTTTTAGTTGGATTAATAGATTTTGTATGG192                           PheValProGlyGlyGlyPheLeuValGlyLeuIleAspPheValTrp                              505560                                                                        GGAATAGTTGGCCCTTCTCAATGGGATGCATTTCTAGTACAAATTGAA240                           GlyIleValGlyProSerGlnTrpAspAlaPheLeuValGlnIleGlu                              65707580                                                                      CAATTAATTAATGAAAGAATAGCTGAATTTGCTAGGAATGCTGCTATT288                           GlnLeuIleAsnGluArgIleAlaGluPheAlaArgAsnAlaAlaIle                              859095                                                                        GCTAATTTAGAAGGATTAGGAAACAATTTCAATATATATGTGGAAGCA336                           AlaAsnLeuGluGlyLeuGlyAsnAsnPheAsnIleTyrValGluAla                              100105110                                                                     TTTAAAGAATGGGAAGAAGATCCTAATAATCCAGAAACCAGGACCAGA384                           PheLysGluTrpGluGluAspProAsnAsnProGluThrArgThrArg                              115120125                                                                     GTAATTGATCGCTTTCGTATACTTGATGGGCTACTTGAAAGGGACATT432                           ValIleAspArgPheArgIleLeuAspGlyLeuLeuGluArgAspIle                              130135140                                                                     CCTTCGTTTCGAATTTCTGGATTTGAAGTACCCCTTTTATCCGTTTAT480                           ProSerPheArgIleSerGlyPheGluValProLeuLeuSerValTyr                              145150155160                                                                  GCTCAAGCGGCCAATCTGCATCTAGCTATATTAAGAGATTCTGTAATT528                           AlaGlnAlaAlaAsnLeuHisLeuAlaIleLeuArgAspSerValIle                              165170175                                                                     TTTGGAGAAAGATGGGGATTGACAACGATAAATGTCAATGAAAACTAT576                           PheGlyGluArgTrpGlyLeuThrThrIleAsnValAsnGluAsnTyr                              180185190                                                                     AATAGACTAATTAGGCATATTGATGAATATGCTGATCACTGTGCAAAT624                           AsnArgLeuIleArgHisIleAspGluTyrAlaAspHisCysAlaAsn                              195200205                                                                     ACGTATAATCGGGGATTAAATAATTTACCGAAATCTACGTATCAAGAT672                           ThrTyrAsnArgGlyLeuAsnAsnLeuProLysSerThrTyrGlnAsp                              210215220                                                                     TGGATAACATATAATCGATTACGGAGAGACTTAACATTGACTGTATTA720                           TrpIleThrTyrAsnArgLeuArgArgAspLeuThrLeuThrValLeu                              225230235240                                                                  GATATCGCCGCTTTCTTTCCAAACTATGACAATAGGAGATATCCAATT768                           AspIleAlaAlaPhePheProAsnTyrAspAsnArgArgTyrProIle                              245250255                                                                     CAGCCAGTTGGTCAACTAACAAGGGAAGTTTATACGGACCCATTAATT816                           GlnProValGlyGlnLeuThrArgGluValTyrThrAspProLeuIle                              260265270                                                                     AATTTTAATCCACAGTTACAGTCTGTAGCTCAATTACCTACTTTTAAC864                           AsnPheAsnProGlnLeuGlnSerValAlaGlnLeuProThrPheAsn                              275280285                                                                     GTTATGGAGAGCAGCGCAATTAGAAATCCTCATTTATTTGATATATTG912                           ValMetGluSerSerAlaIleArgAsnProHisLeuPheAspIleLeu                              290295300                                                                     AATAATCTTACAATCTTTACGGATTGGTTTAGTGTTGGACGCAATTTT960                           AsnAsnLeuThrIlePheThrAspTrpPheSerValGlyArgAsnPhe                              305310315320                                                                  TATTGGGGAGGACATCGAGTAATATCTAGCCTTATAGGAGGTGGTAAC1008                          TyrTrpGlyGlyHisArgValIleSerSerLeuIleGlyGlyGlyAsn                              325330335                                                                     ATAACATCTCCTATATATGGAAGAGAGGCGAACCAGGAGCCTCCAAGA1056                          IleThrSerProIleTyrGlyArgGluAlaAsnGlnGluProProArg                              340345350                                                                     TCCTTTACTTTTAATGGACCGGTATTTAGGACTTTATCAAATCCTACT1104                          SerPheThrPheAsnGlyProValPheArgThrLeuSerAsnProThr                              355360365                                                                     TTACGATTATTACAGCAACCTTGGCCAGCGCCACCATTTAATTTACGT1152                          LeuArgLeuLeuGlnGlnProTrpProAlaProProPheAsnLeuArg                              370375380                                                                     GGTGTTGAAGGAGTAGAATTTTCTACACCTACAAATAGCTTTACGTAT1200                          GlyValGluGlyValGluPheSerThrProThrAsnSerPheThrTyr                              385390395400                                                                  CGAGGAAGAGGTACGGTTGATTCTTTAACTGAATTACCGCCTGAGGAT1248                          ArgGlyArgGlyThrValAspSerLeuThrGluLeuProProGluAsp                              405410415                                                                     AATAGTGTGCCACCTCGCGAAGGATATAGTCATCGTTTATGTCATGCA1296                          AsnSerValProProArgGluGlyTyrSerHisArgLeuCysHisAla                              420425430                                                                     ACTTTTGTTCAAAGATCTGGAACACCTTTTTTAACAACTGGTGTAGTA1344                          ThrPheValGlnArgSerGlyThrProPheLeuThrThrGlyValVal                              435440445                                                                     TTTTCTTGGACGCATCGTAGTGCAACTCTTACAAATACAATTGATCCA1392                          PheSerTrpThrHisArgSerAlaThrLeuThrAsnThrIleAspPro                              450455460                                                                     GAGAGAATTAATCAAATACCTTTAGTGAAAGGATTTAGAGTTTGGGGG1440                          GluArgIleAsnGlnIleProLeuValLysGlyPheArgValTrpGly                              465470475480                                                                  GGCACCTCTGTCATTACAGGACCAGGATTTACAGGAGGGGATATCCTT1488                          GlyThrSerValIleThrGlyProGlyPheThrGlyGlyAspIleLeu                              485490495                                                                     CGAAGAAATACCTTTGGTGATTTTGTATCTCTACAAGTCAATATTAAT1536                          ArgArgAsnThrPheGlyAspPheValSerLeuGlnValAsnIleAsn                              500505510                                                                     TCACCAATTACCCAAAGATACCGTTTAAGATTTCGTTACGCTTCCAGT1584                          SerProIleThrGlnArgTyrArgLeuArgPheArgTyrAlaSerSer                              515520525                                                                     AGGGATGCACGAGTTATAGTATTAACAGGAGCGGCATCCACAGGAGTG1632                          ArgAspAlaArgValIleValLeuThrGlyAlaAlaSerThrGlyVal                              530535540                                                                     GGAGGCCAAGTTAGTGTAAATATGCCTCTTCAGAAAACTATGGAAATA1680                          GlyGlyGlnValSerValAsnMetProLeuGlnLysThrMetGluIle                              545550555560                                                                  GGGGAGAACTTAACATCTAGAACATTTAGATATACCGATTTTAGTAAT1728                          GlyGluAsnLeuThrSerArgThrPheArgTyrThrAspPheSerAsn                              565570575                                                                     CCTTTTTCATTTAGAGCTAATCCAGATATAATTGGGATAAGTGAACAA1776                          ProPheSerPheArgAlaAsnProAspIleIleGlyIleSerGluGln                              580585590                                                                     CCTCTATTTGGTGCAGGTTCTATTAGTAGCGGTGAACTTTATATAGAT1824                          ProLeuPheGlyAlaGlySerIleSerSerGlyGluLeuTyrIleAsp                              595600605                                                                     AAAATTGAAATTATTCTAGCAGATGCAACATTTGAAGCAGAATCTGAT1872                          LysIleGluIleIleLeuAlaAspAlaThrPheGluAlaGluSerAsp                              610615620                                                                     TTAGAAAGAGCACAAAAGGCGGTGAATGCCCTGTTTACTTCTTCCAAT1920                          LeuGluArgAlaGlnLysAlaValAsnAlaLeuPheThrSerSerAsn                              625630635640                                                                  CAAATCGGGTTAAAAACCGATGTGACGGATTATCATATTGATCAAGTA1968                          GlnIleGlyLeuLysThrAspValThrAspTyrHisIleAspGlnVal                              645650655                                                                     TCCAATTTAGTGGATTGTTTATCAGATGAATTTTGTCTGGATGAAAAG2016                          SerAsnLeuValAspCysLeuSerAspGluPheCysLeuAspGluLys                              660665670                                                                     CGAGAATTGTCCGAGAAAGTCAAACATGCGAAGCGACTCAGTGATGAG2064                          ArgGluLeuSerGluLysValLysHisAlaLysArgLeuSerAspGlu                              675680685                                                                     CGGAATTTACTTCAAGATCCAAACTTCAGAGGGATCAATAGACAACCA2112                          ArgAsnLeuLeuGlnAspProAsnPheArgGlyIleAsnArgGlnPro                              690695700                                                                     GACCGTGGCTGGAGAGGAAGTACAGATATTACCATCCAAGGAGGAGAT2160                          AspArgGlyTrpArgGlySerThrAspIleThrIleGlnGlyGlyAsp                              705710715720                                                                  GACGTATTCAAAGAGAATTACGTCACACTACCGGGTACCGTTGATGAG2208                          AspValPheLysGluAsnTyrValThrLeuProGlyThrValAspGlu                              725730735                                                                     TGCTATCCAACGTATTTATATCAGAAAATAGATGAGTCGAAATTAAAA2256                          CysTyrProThrTyrLeuTyrGlnLysIleAspGluSerLysLeuLys                              740745750                                                                     GCTTATACCCGTTATGAATTAAGAGGGTATATCGAAGATAGTCAAGAC2304                          AlaTyrThrArgTyrGluLeuArgGlyTyrIleGluAspSerGlnAsp                              755760765                                                                     TTAGAAATCTATTTGATCCGTTACAATGCAAAACACGAAATAGTAAAT2352                          LeuGluIleTyrLeuIleArgTyrAsnAlaLysHisGluIleValAsn                              770775780                                                                     GTGCCAGGCACGGGTTCCTTATGGCCGCTTTCAGCCCAAAGTCCAATC2400                          ValProGlyThrGlySerLeuTrpProLeuSerAlaGlnSerProIle                              785790795800                                                                  GGAAAGTGTGGAGAACCGAATCGATGCGCGCCACACCTTGAATGGAAT2448                          GlyLysCysGlyGluProAsnArgCysAlaProHisLeuGluTrpAsn                              805810815                                                                     CCTGATCTAGATTGTTCCTGCAGAGACGGGGAAAAATGTGCACATCAT2496                          ProAspLeuAspCysSerCysArgAspGlyGluLysCysAlaHisHis                              820825830                                                                     TCCCATCATTTCACCTTGGATATTGATGTTGGATGTACAGACTTAAAT2544                          SerHisHisPheThrLeuAspIleAspValGlyCysThrAspLeuAsn                              835840845                                                                     GAGGACTTAGGTGTATGGGTGATATTCAAGATTAAGACGCAAGATGGC2592                          GluAspLeuGlyValTrpValIlePheLysIleLysThrGlnAspGly                              850855860                                                                     CATGCAAGACTAGGGAATCTAGAGTTTCTCGAAGAGAAACCATTATTA2640                          HisAlaArgLeuGlyAsnLeuGluPheLeuGluGluLysProLeuLeu                              865870875880                                                                  GGGGAAGCACTAGCTCGTGTGAAAAGAGCGGAGAAGAAGTGGAGAGAC2688                          GlyGluAlaLeuAlaArgValLysArgAlaGluLysLysTrpArgAsp                              885890895                                                                     AAACGAGAGAAACTGCAGTTGGAAACAAATATTGTTTATAAAGAGGCA2736                          LysArgGluLysLeuGlnLeuGluThrAsnIleValTyrLysGluAla                              900905910                                                                     AAAGAATCTGTAGATGCTTTATTTGTAAACTCTCAATATGATAGATTA2784                          LysGluSerValAspAlaLeuPheValAsnSerGlnTyrAspArgLeu                              915920925                                                                     CAAGTGGATACGAACATCGCGATGATTCATGCGGCAGATAAACGCGTT2832                          GlnValAspThrAsnIleAlaMetIleHisAlaAlaAspLysArgVal                              930935940                                                                     CATAGAATCCGGGAAGCGTATCTGCCAGAGTTGTCTGTGATTCCAGGT2880                          HisArgIleArgGluAlaTyrLeuProGluLeuSerValIleProGly                              945950955960                                                                  GTCAATGCGGCCATTTTCGAAGAATTAGAGGGACGTATTTTTACAGCG2928                          ValAsnAlaAlaIlePheGluGluLeuGluGlyArgIlePheThrAla                              965970975                                                                     TATTCCTTATATGATGCGAGAAATGTCATTAAAAATGGCGATTTCAAT2976                          TyrSerLeuTyrAspAlaArgAsnValIleLysAsnGlyAspPheAsn                              980985990                                                                     AATGGCTTATTATGCTGGAACGTGAAAGGTCATGTAGATGTAGAAGAG3024                          AsnGlyLeuLeuCysTrpAsnValLysGlyHisValAspValGluGlu                              99510001005                                                                   CAAAACAACCACCGTTCGGTCCTTGTTATCCCAGAATGGGAGGCAGAA3072                          GlnAsnAsnHisArgSerValLeuValIleProGluTrpGluAlaGlu                              101010151020                                                                  GTGTCACAAGAGGTTCGTGTCTGTCCAGGTCGTGGCTATATCCTTCGT3120                          ValSerGlnGluValArgValCysProGlyArgGlyTyrIleLeuArg                              1025103010351040                                                              GTCACAGCATATAAAGAGGGATATGGAGAGGGCTGCGTAACGATCCAT3168                          ValThrAlaTyrLysGluGlyTyrGlyGluGlyCysValThrIleHis                              104510501055                                                                  GAGATCGAAGACAATACAGACGAACTGAAATTCAGCAACTGTGTAGAA3216                          GluIleGluAspAsnThrAspGluLeuLysPheSerAsnCysValGlu                              106010651070                                                                  GAGGAAGTATATCCAAACAACACAGTAACGTGTAATAATTATACTGGG3264                          GluGluValTyrProAsnAsnThrValThrCysAsnAsnTyrThrGly                              107510801085                                                                  ACTCAAGAAGAATATGAGGGTACGTACACTTCTCGTAATCAAGGATAT3312                          ThrGlnGluGluTyrGluGlyThrTyrThrSerArgAsnGlnGlyTyr                              109010951100                                                                  GACGAAGCCTATGGTAATAACCCTTCCGTACCAGCTGATTACGCTTCA3360                          AspGluAlaTyrGlyAsnAsnProSerValProAlaAspTyrAlaSer                              1105111011151120                                                              GTCTATGAAGAAAAATCGTATACAGATGGACGAAGAGAGAATCCTTGT3408                          ValTyrGluGluLysSerTyrThrAspGlyArgArgGluAsnProCys                              112511301135                                                                  GAATCTAACAGAGGCTATGGGGATTACACACCACTACCGGCTGGTTAT3456                          GluSerAsnArgGlyTyrGlyAspTyrThrProLeuProAlaGlyTyr                              114011451150                                                                  GTAACAAAGGATTTAGAGTACTTCCCAGAGACCGATAAGGTATGGATT3504                          ValThrLysAspLeuGluTyrPheProGluThrAspLysValTrpIle                              115511601165                                                                  GAGATCGGAGAAACAGAAGGAACATTCATCGTGGATAGCGTGGAATTA3552                          GluIleGlyGluThrGluGlyThrPheIleValAspSerValGluLeu                              117011751180                                                                  CTCCTTATGGAGGAA3567                                                           LeuLeuMetGluGlu                                                               1185                                                                          (2) INFORMATION FOR SEQ ID NO: 2:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1189 amino acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:                                      MetGluGluAsnAsnGlnAsnGlnCysIleProTyrAsnCysLeuSer                              151015                                                                        AsnProGluGluValLeuLeuAspGlyGluArgIleSerThrGlyAsn                              202530                                                                        SerSerIleAspIleSerLeuSerLeuValGlnPheLeuValSerAsn                              354045                                                                        PheValProGlyGlyGlyPheLeuValGlyLeuIleAspPheValTrp                              505560                                                                        GlyIleValGlyProSerGlnTrpAspAlaPheLeuValGlnIleGlu                              65707580                                                                      GlnLeuIleAsnGluArgIleAlaGluPheAlaArgAsnAlaAlaIle                              859095                                                                        AlaAsnLeuGluGlyLeuGlyAsnAsnPheAsnIleTyrValGluAla                              100105110                                                                     PheLysGluTrpGluGluAspProAsnAsnProGluThrArgThrArg                              115120125                                                                     ValIleAspArgPheArgIleLeuAspGlyLeuLeuGluArgAspIle                              130135140                                                                     ProSerPheArgIleSerGlyPheGluValProLeuLeuSerValTyr                              145150155160                                                                  AlaGlnAlaAlaAsnLeuHisLeuAlaIleLeuArgAspSerValIle                              165170175                                                                     PheGlyGluArgTrpGlyLeuThrThrIleAsnValAsnGluAsnTyr                              180185190                                                                     AsnArgLeuIleArgHisIleAspGluTyrAlaAspHisCysAlaAsn                              195200205                                                                     ThrTyrAsnArgGlyLeuAsnAsnLeuProLysSerThrTyrGlnAsp                              210215220                                                                     TrpIleThrTyrAsnArgLeuArgArgAspLeuThrLeuThrValLeu                              225230235240                                                                  AspIleAlaAlaPhePheProAsnTyrAspAsnArgArgTyrProIle                              245250255                                                                     GlnProValGlyGlnLeuThrArgGluValTyrThrAspProLeuIle                              260265270                                                                     AsnPheAsnProGlnLeuGlnSerValAlaGlnLeuProThrPheAsn                              275280285                                                                     ValMetGluSerSerAlaIleArgAsnProHisLeuPheAspIleLeu                              290295300                                                                     AsnAsnLeuThrIlePheThrAspTrpPheSerValGlyArgAsnPhe                              305310315320                                                                  TyrTrpGlyGlyHisArgValIleSerSerLeuIleGlyGlyGlyAsn                              325330335                                                                     IleThrSerProIleTyrGlyArgGluAlaAsnGlnGluProProArg                              340345350                                                                     SerPheThrPheAsnGlyProValPheArgThrLeuSerAsnProThr                              355360365                                                                     LeuArgLeuLeuGlnGlnProTrpProAlaProProPheAsnLeuArg                              370375380                                                                     GlyValGluGlyValGluPheSerThrProThrAsnSerPheThrTyr                              385390395400                                                                  ArgGlyArgGlyThrValAspSerLeuThrGluLeuProProGluAsp                              405410415                                                                     AsnSerValProProArgGluGlyTyrSerHisArgLeuCysHisAla                              420425430                                                                     ThrPheValGlnArgSerGlyThrProPheLeuThrThrGlyValVal                              435440445                                                                     PheSerTrpThrHisArgSerAlaThrLeuThrAsnThrIleAspPro                              450455460                                                                     GluArgIleAsnGlnIleProLeuValLysGlyPheArgValTrpGly                              465470475480                                                                  GlyThrSerValIleThrGlyProGlyPheThrGlyGlyAspIleLeu                              485490495                                                                     ArgArgAsnThrPheGlyAspPheValSerLeuGlnValAsnIleAsn                              500505510                                                                     SerProIleThrGlnArgTyrArgLeuArgPheArgTyrAlaSerSer                              515520525                                                                     ArgAspAlaArgValIleValLeuThrGlyAlaAlaSerThrGlyVal                              530535540                                                                     GlyGlyGlnValSerValAsnMetProLeuGlnLysThrMetGluIle                              545550555560                                                                  GlyGluAsnLeuThrSerArgThrPheArgTyrThrAspPheSerAsn                              565570575                                                                     ProPheSerPheArgAlaAsnProAspIleIleGlyIleSerGluGln                              580585590                                                                     ProLeuPheGlyAlaGlySerIleSerSerGlyGluLeuTyrIleAsp                              595600605                                                                     LysIleGluIleIleLeuAlaAspAlaThrPheGluAlaGluSerAsp                              610615620                                                                     LeuGluArgAlaGlnLysAlaValAsnAlaLeuPheThrSerSerAsn                              625630635640                                                                  GlnIleGlyLeuLysThrAspValThrAspTyrHisIleAspGlnVal                              645650655                                                                     SerAsnLeuValAspCysLeuSerAspGluPheCysLeuAspGluLys                              660665670                                                                     ArgGluLeuSerGluLysValLysHisAlaLysArgLeuSerAspGlu                              675680685                                                                     ArgAsnLeuLeuGlnAspProAsnPheArgGlyIleAsnArgGlnPro                              690695700                                                                     AspArgGlyTrpArgGlySerThrAspIleThrIleGlnGlyGlyAsp                              705710715720                                                                  AspValPheLysGluAsnTyrValThrLeuProGlyThrValAspGlu                              725730735                                                                     CysTyrProThrTyrLeuTyrGlnLysIleAspGluSerLysLeuLys                              740745750                                                                     AlaTyrThrArgTyrGluLeuArgGlyTyrIleGluAspSerGlnAsp                              755760765                                                                     LeuGluIleTyrLeuIleArgTyrAsnAlaLysHisGluIleValAsn                              770775780                                                                     ValProGlyThrGlySerLeuTrpProLeuSerAlaGlnSerProIle                              785790795800                                                                  GlyLysCysGlyGluProAsnArgCysAlaProHisLeuGluTrpAsn                              805810815                                                                     ProAspLeuAspCysSerCysArgAspGlyGluLysCysAlaHisHis                              820825830                                                                     SerHisHisPheThrLeuAspIleAspValGlyCysThrAspLeuAsn                              835840845                                                                     GluAspLeuGlyValTrpValIlePheLysIleLysThrGlnAspGly                              850855860                                                                     HisAlaArgLeuGlyAsnLeuGluPheLeuGluGluLysProLeuLeu                              865870875880                                                                  GlyGluAlaLeuAlaArgValLysArgAlaGluLysLysTrpArgAsp                              885890895                                                                     LysArgGluLysLeuGlnLeuGluThrAsnIleValTyrLysGluAla                              900905910                                                                     LysGluSerValAspAlaLeuPheValAsnSerGlnTyrAspArgLeu                              915920925                                                                     GlnValAspThrAsnIleAlaMetIleHisAlaAlaAspLysArgVal                              930935940                                                                     HisArgIleArgGluAlaTyrLeuProGluLeuSerValIleProGly                              945950955960                                                                  ValAsnAlaAlaIlePheGluGluLeuGluGlyArgIlePheThrAla                              965970975                                                                     TyrSerLeuTyrAspAlaArgAsnValIleLysAsnGlyAspPheAsn                              980985990                                                                     AsnGlyLeuLeuCysTrpAsnValLysGlyHisValAspValGluGlu                              99510001005                                                                   GlnAsnAsnHisArgSerValLeuValIleProGluTrpGluAlaGlu                              101010151020                                                                  ValSerGlnGluValArgValCysProGlyArgGlyTyrIleLeuArg                              1025103010351040                                                              ValThrAlaTyrLysGluGlyTyrGlyGluGlyCysValThrIleHis                              104510501055                                                                  GluIleGluAspAsnThrAspGluLeuLysPheSerAsnCysValGlu                              106010651070                                                                  GluGluValTyrProAsnAsnThrValThrCysAsnAsnTyrThrGly                              107510801085                                                                  ThrGlnGluGluTyrGluGlyThrTyrThrSerArgAsnGlnGlyTyr                              109010951100                                                                  AspGluAlaTyrGlyAsnAsnProSerValProAlaAspTyrAlaSer                              1105111011151120                                                              ValTyrGluGluLysSerTyrThrAspGlyArgArgGluAsnProCys                              112511301135                                                                  GluSerAsnArgGlyTyrGlyAspTyrThrProLeuProAlaGlyTyr                              114011451150                                                                  ValThrLysAspLeuGluTyrPheProGluThrAspLysValTrpIle                              115511601165                                                                  GluIleGlyGluThrGluGlyThrPheIleValAspSerValGluLeu                              117011751180                                                                  LeuLeuMetGluGlu                                                               1185                                                                          (2) INFORMATION FOR SEQ ID NO: 3:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 3513 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: unknown                                                         (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iii) ANTI-SENSE: NO                                                          (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Bacillus thuringiensis                                          (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..3513                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:                                      ATGGAGATAGTGAATAATCAGAATCAATGCGTGCCTTATAATTGTTTA48                            MetGluIleValAsnAsnGlnAsnGlnCysValProTyrAsnCysLeu                              151015                                                                        AATAATCCTGAAAATGAGATATTAGATATTGAAAGGTCAAATAGTACT96                            AsnAsnProGluAsnGluIleLeuAspIleGluArgSerAsnSerThr                              202530                                                                        GTAGCAACAAACATCGCCTTGGAGATTAGTCGTCTGCTCGCTTCCGCA144                           ValAlaThrAsnIleAlaLeuGluIleSerArgLeuLeuAlaSerAla                              354045                                                                        ACTCCAATAGGGGGGATTTTATTAGGATTGTTTGATGCAATATGGGGG192                           ThrProIleGlyGlyIleLeuLeuGlyLeuPheAspAlaIleTrpGly                              505560                                                                        TCTATAGGCCCTTCACAATGGGATTTATTTTTAGAGCAAATTGAGCTA240                           SerIleGlyProSerGlnTrpAspLeuPheLeuGluGlnIleGluLeu                              65707580                                                                      TTGATTGACCAAAAAATAGAGGAATTCGCTAGAAACCAGGCAATTTCT288                           LeuIleAspGlnLysIleGluGluPheAlaArgAsnGlnAlaIleSer                              859095                                                                        AGATTGGAAGGGATAAGCAGTCTGTACGGAATTTATACAGAAGCTTTT336                           ArgLeuGluGlyIleSerSerLeuTyrGlyIleTyrThrGluAlaPhe                              100105110                                                                     AGAGAGTGGGAAGCAGATCCTACTAATCCAGCATTAAAAGAAGAGATG384                           ArgGluTrpGluAlaAspProThrAsnProAlaLeuLysGluGluMet                              115120125                                                                     CGTACTCAATTTAATGACATGAACAGTATTCTTGTAACAGCTATTCCT432                           ArgThrGlnPheAsnAspMetAsnSerIleLeuValThrAlaIlePro                              130135140                                                                     CTTTTTTCAGTTCAAAATTATCAAGTCCCATTTTTATCAGTATATGTT480                           LeuPheSerValGlnAsnTyrGlnValProPheLeuSerValTyrVal                              145150155160                                                                  CAAGCTGCAAATTTACATTTATCGGTTTTGAGAGATGTTTCAGTGTTT528                           GlnAlaAlaAsnLeuHisLeuSerValLeuArgAspValSerValPhe                              165170175                                                                     GGGCAGGCTTGGGGATTTGATATAGCAACAATAAATAGTCGTTATAAT576                           GlyGlnAlaTrpGlyPheAspIleAlaThrIleAsnSerArgTyrAsn                              180185190                                                                     GATCTGACTAGACTTATTCCTATATATACAGATTATGCTGTACGCTGG624                           AspLeuThrArgLeuIleProIleTyrThrAspTyrAlaValArgTrp                              195200205                                                                     TACAATACGGGATTAGATCGCTTACCACGAACTGGTGGGCTGCGAAAC672                           TyrAsnThrGlyLeuAspArgLeuProArgThrGlyGlyLeuArgAsn                              210215220                                                                     TGGGCAAGATTTAATCAGTTTAGAAGAGAGTTAACAATATCAGTATTA720                           TrpAlaArgPheAsnGlnPheArgArgGluLeuThrIleSerValLeu                              225230235240                                                                  GATATTATTTCTTTTTTCAGAAATTACGATTCTAGATTATATCCAATT768                           AspIleIleSerPhePheArgAsnTyrAspSerArgLeuTyrProIle                              245250255                                                                     CCAACAAGCTCCCAATTAACGCGGGAAGTATATACAGATCCGGTAATT816                           ProThrSerSerGlnLeuThrArgGluValTyrThrAspProValIle                              260265270                                                                     AATATAACTGACTATAGAGTTGGCCCCAGCTTCGAGAATATTGAGAAC864                           AsnIleThrAspTyrArgValGlyProSerPheGluAsnIleGluAsn                              275280285                                                                     TCAGCCATTAGAAGCCCCCACCTTATGGACTTCTTAAATAATTTGACC912                           SerAlaIleArgSerProHisLeuMetAspPheLeuAsnAsnLeuThr                              290295300                                                                     ATTGATACGGATTTGATTAGAGGTGTTCACTATTGGGCAGGGCATCGT960                           IleAspThrAspLeuIleArgGlyValHisTyrTrpAlaGlyHisArg                              305310315320                                                                  GTAACTTCTCATTTTACAGGTAGTTCTCAAGTGATAACAACCCCTCAA1008                          ValThrSerHisPheThrGlySerSerGlnValIleThrThrProGln                              325330335                                                                     TATGGGATAACCGCAAATGCGGAACCAAGACGAACTATTGCTCCTAGT1056                          TyrGlyIleThrAlaAsnAlaGluProArgArgThrIleAlaProSer                              340345350                                                                     ACTTTTCCAGGTCTTAACCTATTTTATAGAACATTATCAAATCCTTTC1104                          ThrPheProGlyLeuAsnLeuPheTyrArgThrLeuSerAsnProPhe                              355360365                                                                     TTCCGAAGATCAGAAAATATTACTCCTACCTTAGGGATAAATGTAGTA1152                          PheArgArgSerGluAsnIleThrProThrLeuGlyIleAsnValVal                              370375380                                                                     CAGGGAGTAGGGTTCATTCAACCAAATAATGCTGAAGTTCTATATAGA1200                          GlnGlyValGlyPheIleGlnProAsnAsnAlaGluValLeuTyrArg                              385390395400                                                                  AGTAGGGGGACAGTAGATTCTCTTAATGAGTTACCAATTGATGGTGAG1248                          SerArgGlyThrValAspSerLeuAsnGluLeuProIleAspGlyGlu                              405410415                                                                     AATTCATTAGTTGGATATAGTCATCGATTAAGTCATGTTACACTAACC1296                          AsnSerLeuValGlyTyrSerHisArgLeuSerHisValThrLeuThr                              420425430                                                                     AGGTCGTTATATAATACTAATATAACTAGCCTGCCAACATTTGTTTGG1344                          ArgSerLeuTyrAsnThrAsnIleThrSerLeuProThrPheValTrp                              435440445                                                                     ACACATCACAGTGCTACTAATACAAATACAATTAATCCAGATATTATT1392                          ThrHisHisSerAlaThrAsnThrAsnThrIleAsnProAspIleIle                              450455460                                                                     ACACAAATACCTTTAGTGAAAGGATTTAGACTTGGTGGTGGCACCTCT1440                          ThrGlnIleProLeuValLysGlyPheArgLeuGlyGlyGlyThrSer                              465470475480                                                                  GTCATTAAAGGACCAGGATTTACAGGAGGGGATATCCTTCGAAGAAAT1488                          ValIleLysGlyProGlyPheThrGlyGlyAspIleLeuArgArgAsn                              485490495                                                                     ACCATTGGTGAGTTTGTGTCTTTACAAGTCAATATTAACTCACCAATT1536                          ThrIleGlyGluPheValSerLeuGlnValAsnIleAsnSerProIle                              500505510                                                                     ACCCAAAGATACCGTTTAAGATTTCGTTATGCTTCCAGTAGGGATGCA1584                          ThrGlnArgTyrArgLeuArgPheArgTyrAlaSerSerArgAspAla                              515520525                                                                     CGAATTACTGTAGCGATAGGAGGACAAATTAGAGTAGATATGACCCTT1632                          ArgIleThrValAlaIleGlyGlyGlnIleArgValAspMetThrLeu                              530535540                                                                     GAAAAAACCATGGAAATTGGGGAGAGCTTAACATCTAGAACATTTAGC1680                          GluLysThrMetGluIleGlyGluSerLeuThrSerArgThrPheSer                              545550555560                                                                  TATACCAATTTTAGTAATCCTTTTTCATTTAGGGCTAATCCAGATATA1728                          TyrThrAsnPheSerAsnProPheSerPheArgAlaAsnProAspIle                              565570575                                                                     ATTAGAATAGCTGAAGAACTTCCTATTCGTGGTGGTGAGCTTTATATA1776                          IleArgIleAlaGluGluLeuProIleArgGlyGlyGluLeuTyrIle                              580585590                                                                     GATAAAATTGAACTTATTCTAGCAGATGCAACATTTGAAGAAGAATAT1824                          AspLysIleGluLeuIleLeuAlaAspAlaThrPheGluGluGluTyr                              595600605                                                                     GATTTGGAAAGAGCACAGAAGGCGGTGAATGCCCTGTTTACTTCTACA1872                          AspLeuGluArgAlaGlnLysAlaValAsnAlaLeuPheThrSerThr                              610615620                                                                     AATCAACTAGGGCTAAAAACAGATGTGACGGATTATCATATTGATCAA1920                          AsnGlnLeuGlyLeuLysThrAspValThrAspTyrHisIleAspGln                              625630635640                                                                  GTTTCCAATTTAGTTGAGTGTTTATCGGATGAATTTTGTCTGGATGAA1968                          ValSerAsnLeuValGluCysLeuSerAspGluPheCysLeuAspGlu                              645650655                                                                     AAGAGAGAATTATCCGAGAAAGTCAAACATGCGAAGCGACTCAGTGAT2016                          LysArgGluLeuSerGluLysValLysHisAlaLysArgLeuSerAsp                              660665670                                                                     GAACGGAATTTACTTCAAGATCCAAACTTCAGAGGGATCAATAGGCAA2064                          GluArgAsnLeuLeuGlnAspProAsnPheArgGlyIleAsnArgGln                              675680685                                                                     CCAGACCGTGGCTGGAGAGGAAGCACGGATATTACTATCCAAGGTGGA2112                          ProAspArgGlyTrpArgGlySerThrAspIleThrIleGlnGlyGly                              690695700                                                                     GATGACGTATTCAAAGAGAATTACGTCACATTACCGGGTACCTTTGAT2160                          AspAspValPheLysGluAsnTyrValThrLeuProGlyThrPheAsp                              705710715720                                                                  GAGTGCTATCCAACGTATTTATATCAAAAAATAGATGAGTCGAAGTTA2208                          GluCysTyrProThrTyrLeuTyrGlnLysIleAspGluSerLysLeu                              725730735                                                                     AAAGCTTATACCCGCTATGAATTAAGAGGGTATATCGAGGATAGTCAA2256                          LysAlaTyrThrArgTyrGluLeuArgGlyTyrIleGluAspSerGln                              740745750                                                                     GACTTAGAAATCTATTTAATTCGCTACAATGCAAAACACGAGACAGTA2304                          AspLeuGluIleTyrLeuIleArgTyrAsnAlaLysHisGluThrVal                              755760765                                                                     AACGTGCCAGGTACGGGTTCCTTATGGCCGCTTTCAGCCCAAAGTCCA2352                          AsnValProGlyThrGlySerLeuTrpProLeuSerAlaGlnSerPro                              770775780                                                                     ATCGGAAAGTGTGGAGAACCGAATCGATGCGCGCCACACCTTGAATGG2400                          IleGlyLysCysGlyGluProAsnArgCysAlaProHisLeuGluTrp                              785790795800                                                                  AATCCTAATCTAGATTGCTCCTGCAGAGACGGGGAAAAATGTGCCCAT2448                          AsnProAsnLeuAspCysSerCysArgAspGlyGluLysCysAlaHis                              805810815                                                                     CATTCCCATCATTTCTCCTTGGACATTGATGTTGGATGTACAGACTTA2496                          HisSerHisHisPheSerLeuAspIleAspValGlyCysThrAspLeu                              820825830                                                                     AATGAGGACTTAGGTGTATGGGTGATATTCAAGATTAAGACACAAGAT2544                          AsnGluAspLeuGlyValTrpValIlePheLysIleLysThrGlnAsp                              835840845                                                                     GGCTATGCAAGACTAGGAAATCTAGAGTTTCTCGAAGAGAACCCACTA2592                          GlyTyrAlaArgLeuGlyAsnLeuGluPheLeuGluGluAsnProLeu                              850855860                                                                     TTAGGGGAAGCACTAGCTCGTGTGAAAAGAGCGGAGAAAAAATGGAGA2640                          LeuGlyGluAlaLeuAlaArgValLysArgAlaGluLysLysTrpArg                              865870875880                                                                  GACAAATGCGAAAAATTGGAATGGGAAACAAATATTGTTTATAAAGAG2688                          AspLysCysGluLysLeuGluTrpGluThrAsnIleValTyrLysGlu                              885890895                                                                     GCAAAAGAATCTGTAGATGCTTTATTTGTAAACTCTCAATATGATAGA2736                          AlaLysGluSerValAspAlaLeuPheValAsnSerGlnTyrAspArg                              900905910                                                                     TTACAAGCGGATACGAATATCGCGATGATTCATGCGGCAGATAAACGC2784                          LeuGlnAlaAspThrAsnIleAlaMetIleHisAlaAlaAspLysArg                              915920925                                                                     GTTCATAGCATTCGAGAAGCGTATCTGCCAGAGCTGTCTGTGATTCCG2832                          ValHisSerIleArgGluAlaTyrLeuProGluLeuSerValIlePro                              930935940                                                                     GGTGTCAATGCGGCTATTTTTGAAGAATTAGAAGGGCGTATTTTCACT2880                          GlyValAsnAlaAlaIlePheGluGluLeuGluGlyArgIlePheThr                              945950955960                                                                  GCATTCTCCCTATATGATGCGAGAAATGTCATTAAAAATGGCGATTTC2928                          AlaPheSerLeuTyrAspAlaArgAsnValIleLysAsnGlyAspPhe                              965970975                                                                     AATAATGGCTTATCATGCTGGAACGTGAAAGGGCATGTAGATGTAGAA2976                          AsnAsnGlyLeuSerCysTrpAsnValLysGlyHisValAspValGlu                              980985990                                                                     GAACAGAACAACCATCGTTCGGTCCTTGTTGTTCCAGAATGGGAAGCA3024                          GluGlnAsnAsnHisArgSerValLeuValValProGluTrpGluAla                              99510001005                                                                   GAAGTGTCACAAGAAGTTCGTGTTTGTCCGGGTCGTGGCTATATCCTT3072                          GluValSerGlnGluValArgValCysProGlyArgGlyTyrIleLeu                              101010151020                                                                  CGTGTTACAGCGTACAAAGAGGGATATGGAGAGGGCTGTGTAACGATT3120                          ArgValThrAlaTyrLysGluGlyTyrGlyGluGlyCysValThrIle                              1025103010351040                                                              CATGAGATCGAAGACAATACAGACGAACTGAAATTCAGCAACTGTGTA3168                          HisGluIleGluAspAsnThrAspGluLeuLysPheSerAsnCysVal                              104510501055                                                                  GAAGAGGAAGTATATCCAAACAACACGGTAACGTGTAATAATTATACT3216                          GluGluGluValTyrProAsnAsnThrValThrCysAsnAsnTyrThr                              106010651070                                                                  GCGACTCAAGAAGAACATGAGGGTACGTACACTTCCCGTAATCGAGGA3264                          AlaThrGlnGluGluHisGluGlyThrTyrThrSerArgAsnArgGly                              107510801085                                                                  TATGACGAAGCCTATGAAAGCAATTCTTCTGTACATGCGTCAGTCTAT3312                          TyrAspGluAlaTyrGluSerAsnSerSerValHisAlaSerValTyr                              109010951100                                                                  GAAGAAAAATCGTATACAGATAGACGAAGAGAGAATCCTTGTGAATCT3360                          GluGluLysSerTyrThrAspArgArgArgGluAsnProCysGluSer                              1105111011151120                                                              AACAGAGGATATGGGGATTACACACCACTACCAGCTGGCTATGTGACA3408                          AsnArgGlyTyrGlyAspTyrThrProLeuProAlaGlyTyrValThr                              112511301135                                                                  AAAGAATTAGAGTACTTCCCAGAAACCGATAAGGTATGGATTGAGATC3456                          LysGluLeuGluTyrPheProGluThrAspLysValTrpIleGluIle                              114011451150                                                                  GGAGAAACGGAAGGAACATTCATCGTGGACAGCGTGGAATTACTTCTT3504                          GlyGluThrGluGlyThrPheIleValAspSerValGluLeuLeuLeu                              115511601165                                                                  ATGGAGGAA3513                                                                 MetGluGlu                                                                     1170                                                                          (2) INFORMATION FOR SEQ ID NO: 4:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1171 amino acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:                                      MetGluIleValAsnAsnGlnAsnGlnCysValProTyrAsnCysLeu                              151015                                                                        AsnAsnProGluAsnGluIleLeuAspIleGluArgSerAsnSerThr                              202530                                                                        ValAlaThrAsnIleAlaLeuGluIleSerArgLeuLeuAlaSerAla                              354045                                                                        ThrProIleGlyGlyIleLeuLeuGlyLeuPheAspAlaIleTrpGly                              505560                                                                        SerIleGlyProSerGlnTrpAspLeuPheLeuGluGlnIleGluLeu                              65707580                                                                      LeuIleAspGlnLysIleGluGluPheAlaArgAsnGlnAlaIleSer                              859095                                                                        ArgLeuGluGlyIleSerSerLeuTyrGlyIleTyrThrGluAlaPhe                              100105110                                                                     ArgGluTrpGluAlaAspProThrAsnProAlaLeuLysGluGluMet                              115120125                                                                     ArgThrGlnPheAsnAspMetAsnSerIleLeuValThrAlaIlePro                              130135140                                                                     LeuPheSerValGlnAsnTyrGlnValProPheLeuSerValTyrVal                              145150155160                                                                  GlnAlaAlaAsnLeuHisLeuSerValLeuArgAspValSerValPhe                              165170175                                                                     GlyGlnAlaTrpGlyPheAspIleAlaThrIleAsnSerArgTyrAsn                              180185190                                                                     AspLeuThrArgLeuIleProIleTyrThrAspTyrAlaValArgTrp                              195200205                                                                     TyrAsnThrGlyLeuAspArgLeuProArgThrGlyGlyLeuArgAsn                              210215220                                                                     TrpAlaArgPheAsnGlnPheArgArgGluLeuThrIleSerValLeu                              225230235240                                                                  AspIleIleSerPhePheArgAsnTyrAspSerArgLeuTyrProIle                              245250255                                                                     ProThrSerSerGlnLeuThrArgGluValTyrThrAspProValIle                              260265270                                                                     AsnIleThrAspTyrArgValGlyProSerPheGluAsnIleGluAsn                              275280285                                                                     SerAlaIleArgSerProHisLeuMetAspPheLeuAsnAsnLeuThr                              290295300                                                                     IleAspThrAspLeuIleArgGlyValHisTyrTrpAlaGlyHisArg                              305310315320                                                                  ValThrSerHisPheThrGlySerSerGlnValIleThrThrProGln                              325330335                                                                     TyrGlyIleThrAlaAsnAlaGluProArgArgThrIleAlaProSer                              340345350                                                                     ThrPheProGlyLeuAsnLeuPheTyrArgThrLeuSerAsnProPhe                              355360365                                                                     PheArgArgSerGluAsnIleThrProThrLeuGlyIleAsnValVal                              370375380                                                                     GlnGlyValGlyPheIleGlnProAsnAsnAlaGluValLeuTyrArg                              385390395400                                                                  SerArgGlyThrValAspSerLeuAsnGluLeuProIleAspGlyGlu                              405410415                                                                     AsnSerLeuValGlyTyrSerHisArgLeuSerHisValThrLeuThr                              420425430                                                                     ArgSerLeuTyrAsnThrAsnIleThrSerLeuProThrPheValTrp                              435440445                                                                     ThrHisHisSerAlaThrAsnThrAsnThrIleAsnProAspIleIle                              450455460                                                                     ThrGlnIleProLeuValLysGlyPheArgLeuGlyGlyGlyThrSer                              465470475480                                                                  ValIleLysGlyProGlyPheThrGlyGlyAspIleLeuArgArgAsn                              485490495                                                                     ThrIleGlyGluPheValSerLeuGlnValAsnIleAsnSerProIle                              500505510                                                                     ThrGlnArgTyrArgLeuArgPheArgTyrAlaSerSerArgAspAla                              515520525                                                                     ArgIleThrValAlaIleGlyGlyGlnIleArgValAspMetThrLeu                              530535540                                                                     GluLysThrMetGluIleGlyGluSerLeuThrSerArgThrPheSer                              545550555560                                                                  TyrThrAsnPheSerAsnProPheSerPheArgAlaAsnProAspIle                              565570575                                                                     IleArgIleAlaGluGluLeuProIleArgGlyGlyGluLeuTyrIle                              580585590                                                                     AspLysIleGluLeuIleLeuAlaAspAlaThrPheGluGluGluTyr                              595600605                                                                     AspLeuGluArgAlaGlnLysAlaValAsnAlaLeuPheThrSerThr                              610615620                                                                     AsnGlnLeuGlyLeuLysThrAspValThrAspTyrHisIleAspGln                              625630635640                                                                  ValSerAsnLeuValGluCysLeuSerAspGluPheCysLeuAspGlu                              645650655                                                                     LysArgGluLeuSerGluLysValLysHisAlaLysArgLeuSerAsp                              660665670                                                                     GluArgAsnLeuLeuGlnAspProAsnPheArgGlyIleAsnArgGln                              675680685                                                                     ProAspArgGlyTrpArgGlySerThrAspIleThrIleGlnGlyGly                              690695700                                                                     AspAspValPheLysGluAsnTyrValThrLeuProGlyThrPheAsp                              705710715720                                                                  GluCysTyrProThrTyrLeuTyrGlnLysIleAspGluSerLysLeu                              725730735                                                                     LysAlaTyrThrArgTyrGluLeuArgGlyTyrIleGluAspSerGln                              740745750                                                                     AspLeuGluIleTyrLeuIleArgTyrAsnAlaLysHisGluThrVal                              755760765                                                                     AsnValProGlyThrGlySerLeuTrpProLeuSerAlaGlnSerPro                              770775780                                                                     IleGlyLysCysGlyGluProAsnArgCysAlaProHisLeuGluTrp                              785790795800                                                                  AsnProAsnLeuAspCysSerCysArgAspGlyGluLysCysAlaHis                              805810815                                                                     HisSerHisHisPheSerLeuAspIleAspValGlyCysThrAspLeu                              820825830                                                                     AsnGluAspLeuGlyValTrpValIlePheLysIleLysThrGlnAsp                              835840845                                                                     GlyTyrAlaArgLeuGlyAsnLeuGluPheLeuGluGluAsnProLeu                              850855860                                                                     LeuGlyGluAlaLeuAlaArgValLysArgAlaGluLysLysTrpArg                              865870875880                                                                  AspLysCysGluLysLeuGluTrpGluThrAsnIleValTyrLysGlu                              885890895                                                                     AlaLysGluSerValAspAlaLeuPheValAsnSerGlnTyrAspArg                              900905910                                                                     LeuGlnAlaAspThrAsnIleAlaMetIleHisAlaAlaAspLysArg                              915920925                                                                     ValHisSerIleArgGluAlaTyrLeuProGluLeuSerValIlePro                              930935940                                                                     GlyValAsnAlaAlaIlePheGluGluLeuGluGlyArgIlePheThr                              945950955960                                                                  AlaPheSerLeuTyrAspAlaArgAsnValIleLysAsnGlyAspPhe                              965970975                                                                     AsnAsnGlyLeuSerCysTrpAsnValLysGlyHisValAspValGlu                              980985990                                                                     GluGlnAsnAsnHisArgSerValLeuValValProGluTrpGluAla                              99510001005                                                                   GluValSerGlnGluValArgValCysProGlyArgGlyTyrIleLeu                              101010151020                                                                  ArgValThrAlaTyrLysGluGlyTyrGlyGluGlyCysValThrIle                              1025103010351040                                                              HisGluIleGluAspAsnThrAspGluLeuLysPheSerAsnCysVal                              104510501055                                                                  GluGluGluValTyrProAsnAsnThrValThrCysAsnAsnTyrThr                              106010651070                                                                  AlaThrGlnGluGluHisGluGlyThrTyrThrSerArgAsnArgGly                              107510801085                                                                  TyrAspGluAlaTyrGluSerAsnSerSerValHisAlaSerValTyr                              109010951100                                                                  GluGluLysSerTyrThrAspArgArgArgGluAsnProCysGluSer                              1105111011151120                                                              AsnArgGlyTyrGlyAspTyrThrProLeuProAlaGlyTyrValThr                              112511301135                                                                  LysGluLeuGluTyrPheProGluThrAspLysValTrpIleGluIle                              114011451150                                                                  GlyGluThrGluGlyThrPheIleValAspSerValGluLeuLeuLeu                              115511601165                                                                  MetGluGlu                                                                     1170                                                                          (2) INFORMATION FOR SEQ ID NO: 5:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 3558 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: unknown                                                         (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iii) ANTI-SENSE: NO                                                          (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Hybrid sequence                                                 (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..3558                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:                                      ATGGAGATAGTGAATAATCAGAATCAATGCGTGCCTTATAATTGTTTA48                            MetGluIleValAsnAsnGlnAsnGlnCysValProTyrAsnCysLeu                              151015                                                                        AATAATCCTGAAAATGAGATATTAGATATTGAAAGGTCAAATAGTACT96                            AsnAsnProGluAsnGluIleLeuAspIleGluArgSerAsnSerThr                              202530                                                                        GTAGCAACAAACATCGCCTTGGAGATTAGTCGTCTGCTCGCTTCCGCA144                           ValAlaThrAsnIleAlaLeuGluIleSerArgLeuLeuAlaSerAla                              354045                                                                        ACTCCAATAGGGGGGATTTTATTAGGATTGTTTGATGCAATATGGGGG192                           ThrProIleGlyGlyIleLeuLeuGlyLeuPheAspAlaIleTrpGly                              505560                                                                        TCTATAGGCCCTTCACAATGGGATTTATTTTTAGAGCAAATTGAGCTA240                           SerIleGlyProSerGlnTrpAspLeuPheLeuGluGlnIleGluLeu                              65707580                                                                      TTGATTGACCAAAAAATAGAGGAATTCGCTAGAAACCAGGCAATTTCT288                           LeuIleAspGlnLysIleGluGluPheAlaArgAsnGlnAlaIleSer                              859095                                                                        AGATTGGAAGGGATAAGCAGTCTGTACGGAATTTATACAGAAGCTTTT336                           ArgLeuGluGlyIleSerSerLeuTyrGlyIleTyrThrGluAlaPhe                              100105110                                                                     AGAGAGTGGGAAGCAGATCCTACTAATCCAGCATTAAAAGAAGAGATG384                           ArgGluTrpGluAlaAspProThrAsnProAlaLeuLysGluGluMet                              115120125                                                                     CGTACTCAATTTAATGACATGAACAGTATTCTTGTAACAGCTATTCCT432                           ArgThrGlnPheAsnAspMetAsnSerIleLeuValThrAlaIlePro                              130135140                                                                     CTTTTTTCAGTTCAAAATTATCAAGTCCCATTTTTATCAGTATATGTT480                           LeuPheSerValGlnAsnTyrGlnValProPheLeuSerValTyrVal                              145150155160                                                                  CAAGCTGCAAATTTACATTTATCGGTTTTGAGAGATGTTTCAGTGTTT528                           GlnAlaAlaAsnLeuHisLeuSerValLeuArgAspValSerValPhe                              165170175                                                                     GGGCAGGCTTGGGGATTTGATATAGCAACAATAAATAGTCGTTATAAT576                           GlyGlnAlaTrpGlyPheAspIleAlaThrIleAsnSerArgTyrAsn                              180185190                                                                     GATCTGACTAGACTTATTCCTATATATACAGATTATGCTGTACGCTGG624                           AspLeuThrArgLeuIleProIleTyrThrAspTyrAlaValArgTrp                              195200205                                                                     TACAATACGGGATTAGATCGCTTACCACGAACTGGTGGGCTGCGAAAC672                           TyrAsnThrGlyLeuAspArgLeuProArgThrGlyGlyLeuArgAsn                              210215220                                                                     TGGGCAAGATTTAATCAGTTTAGAAGAGAGTTAACAATATCAGTATTA720                           TrpAlaArgPheAsnGlnPheArgArgGluLeuThrIleSerValLeu                              225230235240                                                                  GATATTATTTCTTTTTTCAGAAATTACGATTCTAGATTATATCCAATT768                           AspIleIleSerPhePheArgAsnTyrAspSerArgLeuTyrProIle                              245250255                                                                     CCAACAAGCTCCCAATTAACGCGGGAAGTATATACAGATCCGGTAATT816                           ProThrSerSerGlnLeuThrArgGluValTyrThrAspProValIle                              260265270                                                                     AATATAACTGACTATAGAGTTGGCCCCAGCTTCGAGAATATTGAGAAC864                           AsnIleThrAspTyrArgValGlyProSerPheGluAsnIleGluAsn                              275280285                                                                     TCAGCCATTAGAAGCCCCCACCTTATGGACTTCTTAAATAATTTGACC912                           SerAlaIleArgSerProHisLeuMetAspPheLeuAsnAsnLeuThr                              290295300                                                                     ATTGATACGGATTTGATTAGAGGTGTTCACTATTGGGCAGGGCATCGT960                           IleAspThrAspLeuIleArgGlyValHisTyrTrpAlaGlyHisArg                              305310315320                                                                  GTAACTTCTCATTTTACAGGTAGTTCTCAAGTGATAACAACCCCTCAA1008                          ValThrSerHisPheThrGlySerSerGlnValIleThrThrProGln                              325330335                                                                     TATGGGATAACCGCAAATGCGGAACCAAGACGAACTATTGCTCCTAGT1056                          TyrGlyIleThrAlaAsnAlaGluProArgArgThrIleAlaProSer                              340345350                                                                     ACTTTTCCAGGTCTTAACCTATTTTATAGAACATTATCAAATCCTTTC1104                          ThrPheProGlyLeuAsnLeuPheTyrArgThrLeuSerAsnProPhe                              355360365                                                                     TTCCGAAGATCAGAAAATATTACTCCTACCTTAGGGATAAATGTAGTA1152                          PheArgArgSerGluAsnIleThrProThrLeuGlyIleAsnValVal                              370375380                                                                     CAGGGAGTAGGGTTCATTCAACCAAATAATGCTGAAGTTCTATATAGA1200                          GlnGlyValGlyPheIleGlnProAsnAsnAlaGluValLeuTyrArg                              385390395400                                                                  AGTAGGGGGACAGTAGATTCTCTTAATGAGTTACCAATTGATGGTGAG1248                          SerArgGlyThrValAspSerLeuAsnGluLeuProIleAspGlyGlu                              405410415                                                                     AATTCATTAGTTGGATATAGTCATCGATTAAGTCATGTTACACTAACC1296                          AsnSerLeuValGlyTyrSerHisArgLeuSerHisValThrLeuThr                              420425430                                                                     AGGTCGTTATATAATACTAATATAACTAGCCTGCCAACATTTGTTTGG1344                          ArgSerLeuTyrAsnThrAsnIleThrSerLeuProThrPheValTrp                              435440445                                                                     ACACATCACAGTGCTACTAATACAAATACAATTAATCCAGATATTATT1392                          ThrHisHisSerAlaThrAsnThrAsnThrIleAsnProAspIleIle                              450455460                                                                     ACACAAATACCTTTAGTGAAAGGATTTAGAGTTTGGGGGGGCACCTCT1440                          ThrGlnIleProLeuValLysGlyPheArgValTrpGlyGlyThrSer                              465470475480                                                                  GTCATTACAGGACCAGGATTTACAGGAGGGGATATCCTTCGAAGAAAT1488                          ValIleThrGlyProGlyPheThrGlyGlyAspIleLeuArgArgAsn                              485490495                                                                     ACCTTTGGTGATTTTGTATCTCTACAAGTCAATATTAATTCACCAATT1536                          ThrPheGlyAspPheValSerLeuGlnValAsnIleAsnSerProIle                              500505510                                                                     ACCCAAAGATACCGTTTAAGATTTCGTTACGCTTCCAGTAGGGATGCA1584                          ThrGlnArgTyrArgLeuArgPheArgTyrAlaSerSerArgAspAla                              515520525                                                                     CGAGTTATAGTATTAACAGGAGCGGCATCCACAGGAGTGGGAGGCCAA1632                          ArgValIleValLeuThrGlyAlaAlaSerThrGlyValGlyGlyGln                              530535540                                                                     GTTAGTGTAAATATGCCTCTTCAGAAAACTATGGAAATAGGGGAGAAC1680                          ValSerValAsnMetProLeuGlnLysThrMetGluIleGlyGluAsn                              545550555560                                                                  TTAACATCTAGAACATTTAGATATACCGATTTTAGTAATCCTTTTTCA1728                          LeuThrSerArgThrPheArgTyrThrAspPheSerAsnProPheSer                              565570575                                                                     TTTAGAGCTAATCCAGATATAATTGGGATAAGTGAACAACCTCTATTT1776                          PheArgAlaAsnProAspIleIleGlyIleSerGluGlnProLeuPhe                              580585590                                                                     GGTGCAGGTTCTATTAGTAGCGGTGAACTTTATATAGATAAAATTGAA1824                          GlyAlaGlySerIleSerSerGlyGluLeuTyrIleAspLysIleGlu                              595600605                                                                     ATTATTCTAGCAGATGCAACATTTGAAGCAGAATCTGATTTAGAAAGA1872                          IleIleLeuAlaAspAlaThrPheGluAlaGluSerAspLeuGluArg                              610615620                                                                     GCACAAAAGGCGGTGAATGCCCTGTTTACTTCTTCCAATCAAATCGGG1920                          AlaGlnLysAlaValAsnAlaLeuPheThrSerSerAsnGlnIleGly                              625630635640                                                                  TTAAAAACCGATGTGACGGATTATCATATTGATCAAGTATCCAATTTA1968                          LeuLysThrAspValThrAspTyrHisIleAspGlnValSerAsnLeu                              645650655                                                                     GTGGATTGTTTATCAGATGAATTTTGTCTGGATGAAAAGCGAGAATTG2016                          ValAspCysLeuSerAspGluPheCysLeuAspGluLysArgGluLeu                              660665670                                                                     TCCGAGAAAGTCAAACATGCGAAGCGACTCAGTGATGAGCGGAATTTA2064                          SerGluLysValLysHisAlaLysArgLeuSerAspGluArgAsnLeu                              675680685                                                                     CTTCAAGATCCAAACTTCAGAGGGATCAATAGACAACCAGACCGTGGC2112                          LeuGlnAspProAsnPheArgGlyIleAsnArgGlnProAspArgGly                              690695700                                                                     TGGAGAGGAAGTACAGATATTACCATCCAAGGAGGAGATGACGTATTC2160                          TrpArgGlySerThrAspIleThrIleGlnGlyGlyAspAspValPhe                              705710715720                                                                  AAAGAGAATTACGTCACACTACCGGGTACCGTTGATGAGTGCTATCCA2208                          LysGluAsnTyrValThrLeuProGlyThrValAspGluCysTyrPro                              725730735                                                                     ACGTATTTATATCAGAAAATAGATGAGTCGAAATTAAAAGCTTATACC2256                          ThrTyrLeuTyrGlnLysIleAspGluSerLysLeuLysAlaTyrThr                              740745750                                                                     CGTTATGAATTAAGAGGGTATATCGAAGATAGTCAAGACTTAGAAATC2304                          ArgTyrGluLeuArgGlyTyrIleGluAspSerGlnAspLeuGluIle                              755760765                                                                     TATTTGATCCGTTACAATGCAAAACACGAAATAGTAAATGTGCCAGGC2352                          TyrLeuIleArgTyrAsnAlaLysHisGluIleValAsnValProGly                              770775780                                                                     ACGGGTTCCTTATGGCCGCTTTCAGCCCAAAGTCCAATCGGAAAGTGT2400                          ThrGlySerLeuTrpProLeuSerAlaGlnSerProIleGlyLysCys                              785790795800                                                                  GGAGAACCGAATCGATGCGCGCCACACCTTGAATGGAATCCTGATCTA2448                          GlyGluProAsnArgCysAlaProHisLeuGluTrpAsnProAspLeu                              805810815                                                                     GATTGTTCCTGCAGAGACGGGGAAAAATGTGCACATCATTCCCATCAT2496                          AspCysSerCysArgAspGlyGluLysCysAlaHisHisSerHisHis                              820825830                                                                     TTCACCTTGGATATTGATGTTGGATGTACAGACTTAAATGAGGACTTA2544                          PheThrLeuAspIleAspValGlyCysThrAspLeuAsnGluAspLeu                              835840845                                                                     GGTGTATGGGTGATATTCAAGATTAAGACGCAAGATGGCCATGCAAGA2592                          GlyValTrpValIlePheLysIleLysThrGlnAspGlyHisAlaArg                              850855860                                                                     CTAGGGAATCTAGAGTTTCTCGAAGAGAAACCATTATTAGGGGAAGCA2640                          LeuGlyAsnLeuGluPheLeuGluGluLysProLeuLeuGlyGluAla                              865870875880                                                                  CTAGCTCGTGTGAAAAGAGCGGAGAAGAAGTGGAGAGACAAACGAGAG2688                          LeuAlaArgValLysArgAlaGluLysLysTrpArgAspLysArgGlu                              885890895                                                                     AAACTGCAGTTGGAAACAAATATTGTTTATAAAGAGGCAAAAGAATCT2736                          LysLeuGlnLeuGluThrAsnIleValTyrLysGluAlaLysGluSer                              900905910                                                                     GTAGATGCTTTATTTGTAAACTCTCAATATGATAGATTACAAGTGGAT2784                          ValAspAlaLeuPheValAsnSerGlnTyrAspArgLeuGlnValAsp                              915920925                                                                     ACGAACATCGCGATGATTCATGCGGCAGATAAACGCGTTCATAGAATC2832                          ThrAsnIleAlaMetIleHisAlaAlaAspLysArgValHisArgIle                              930935940                                                                     CGGGAAGCGTATCTGCCAGAGTTGTCTGTGATTCCAGGTGTCAATGCG2880                          ArgGluAlaTyrLeuProGluLeuSerValIleProGlyValAsnAla                              945950955960                                                                  GCCATTTTCGAAGAATTAGAGGGACGTATTTTTACAGCGTATTCCTTA2928                          AlaIlePheGluGluLeuGluGlyArgIlePheThrAlaTyrSerLeu                              965970975                                                                     TATGATGCGAGAAATGTCATTAAAAATGGCGATTTCAATAATGGCTTA2976                          TyrAspAlaArgAsnValIleLysAsnGlyAspPheAsnAsnGlyLeu                              980985990                                                                     TTATGCTGGAACGTGAAAGGTCATGTAGATGTAGAAGAGCAAAACAAC3024                          LeuCysTrpAsnValLysGlyHisValAspValGluGluGlnAsnAsn                              99510001005                                                                   CACCGTTCGGTCCTTGTTATCCCAGAATGGGAGGCAGAAGTGTCACAA3072                          HisArgSerValLeuValIleProGluTrpGluAlaGluValSerGln                              101010151020                                                                  GAGGTTCGTGTCTGTCCAGGTCGTGGCTATATCCTTCGTGTCACAGCA3120                          GluValArgValCysProGlyArgGlyTyrIleLeuArgValThrAla                              1025103010351040                                                              TATAAAGAGGGATATGGAGAGGGCTGCGTAACGATCCATGAGATCGAA3168                          TyrLysGluGlyTyrGlyGluGlyCysValThrIleHisGluIleGlu                              104510501055                                                                  GACAATACAGACGAACTGAAATTCAGCAACTGTGTAGAAGAGGAAGTA3216                          AspAsnThrAspGluLeuLysPheSerAsnCysValGluGluGluVal                              106010651070                                                                  TATCCAAACAACACAGTAACGTGTAATAATTATACTGGGACTCAAGAA3264                          TyrProAsnAsnThrValThrCysAsnAsnTyrThrGlyThrGlnGlu                              107510801085                                                                  GAATATGAGGGTACGTACACTTCTCGTAATCAAGGATATGACGAAGCC3312                          GluTyrGluGlyThrTyrThrSerArgAsnGlnGlyTyrAspGluAla                              109010951100                                                                  TATGGTAATAACCCTTCCGTACCAGCTGATTACGCTTCAGTCTATGAA3360                          TyrGlyAsnAsnProSerValProAlaAspTyrAlaSerValTyrGlu                              1105111011151120                                                              GAAAAATCGTATACAGATGGACGAAGAGAGAATCCTTGTGAATCTAAC3408                          GluLysSerTyrThrAspGlyArgArgGluAsnProCysGluSerAsn                              112511301135                                                                  AGAGGCTATGGGGATTACACACCACTACCGGCTGGTTATGTAACAAAG3456                          ArgGlyTyrGlyAspTyrThrProLeuProAlaGlyTyrValThrLys                              114011451150                                                                  GATTTAGAGTACTTCCCAGAGACCGATAAGGTATGGATTGAGATCGGA3504                          AspLeuGluTyrPheProGluThrAspLysValTrpIleGluIleGly                              115511601165                                                                  GAAACAGAAGGAACATTCATCGTGGATAGCGTGGAATTACTCCTTATG3552                          GluThrGluGlyThrPheIleValAspSerValGluLeuLeuLeuMet                              117011751180                                                                  GAGGAA3558                                                                    GluGlu                                                                        1185                                                                          (2) INFORMATION FOR SEQ ID NO: 6:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1186 amino acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:                                      MetGluIleValAsnAsnGlnAsnGlnCysValProTyrAsnCysLeu                              151015                                                                        AsnAsnProGluAsnGluIleLeuAspIleGluArgSerAsnSerThr                              202530                                                                        ValAlaThrAsnIleAlaLeuGluIleSerArgLeuLeuAlaSerAla                              354045                                                                        ThrProIleGlyGlyIleLeuLeuGlyLeuPheAspAlaIleTrpGly                              505560                                                                        SerIleGlyProSerGlnTrpAspLeuPheLeuGluGlnIleGluLeu                              65707580                                                                      LeuIleAspGlnLysIleGluGluPheAlaArgAsnGlnAlaIleSer                              859095                                                                        ArgLeuGluGlyIleSerSerLeuTyrGlyIleTyrThrGluAlaPhe                              100105110                                                                     ArgGluTrpGluAlaAspProThrAsnProAlaLeuLysGluGluMet                              115120125                                                                     ArgThrGlnPheAsnAspMetAsnSerIleLeuValThrAlaIlePro                              130135140                                                                     LeuPheSerValGlnAsnTyrGlnValProPheLeuSerValTyrVal                              145150155160                                                                  GlnAlaAlaAsnLeuHisLeuSerValLeuArgAspValSerValPhe                              165170175                                                                     GlyGlnAlaTrpGlyPheAspIleAlaThrIleAsnSerArgTyrAsn                              180185190                                                                     AspLeuThrArgLeuIleProIleTyrThrAspTyrAlaValArgTrp                              195200205                                                                     TyrAsnThrGlyLeuAspArgLeuProArgThrGlyGlyLeuArgAsn                              210215220                                                                     TrpAlaArgPheAsnGlnPheArgArgGluLeuThrIleSerValLeu                              225230235240                                                                  AspIleIleSerPhePheArgAsnTyrAspSerArgLeuTyrProIle                              245250255                                                                     ProThrSerSerGlnLeuThrArgGluValTyrThrAspProValIle                              260265270                                                                     AsnIleThrAspTyrArgValGlyProSerPheGluAsnIleGluAsn                              275280285                                                                     SerAlaIleArgSerProHisLeuMetAspPheLeuAsnAsnLeuThr                              290295300                                                                     IleAspThrAspLeuIleArgGlyValHisTyrTrpAlaGlyHisArg                              305310315320                                                                  ValThrSerHisPheThrGlySerSerGlnValIleThrThrProGln                              325330335                                                                     TyrGlyIleThrAlaAsnAlaGluProArgArgThrIleAlaProSer                              340345350                                                                     ThrPheProGlyLeuAsnLeuPheTyrArgThrLeuSerAsnProPhe                              355360365                                                                     PheArgArgSerGluAsnIleThrProThrLeuGlyIleAsnValVal                              370375380                                                                     GlnGlyValGlyPheIleGlnProAsnAsnAlaGluValLeuTyrArg                              385390395400                                                                  SerArgGlyThrValAspSerLeuAsnGluLeuProIleAspGlyGlu                              405410415                                                                     AsnSerLeuValGlyTyrSerHisArgLeuSerHisValThrLeuThr                              420425430                                                                     ArgSerLeuTyrAsnThrAsnIleThrSerLeuProThrPheValTrp                              435440445                                                                     ThrHisHisSerAlaThrAsnThrAsnThrIleAsnProAspIleIle                              450455460                                                                     ThrGlnIleProLeuValLysGlyPheArgValTrpGlyGlyThrSer                              465470475480                                                                  ValIleThrGlyProGlyPheThrGlyGlyAspIleLeuArgArgAsn                              485490495                                                                     ThrPheGlyAspPheValSerLeuGlnValAsnIleAsnSerProIle                              500505510                                                                     ThrGlnArgTyrArgLeuArgPheArgTyrAlaSerSerArgAspAla                              515520525                                                                     ArgValIleValLeuThrGlyAlaAlaSerThrGlyValGlyGlyGln                              530535540                                                                     ValSerValAsnMetProLeuGlnLysThrMetGluIleGlyGluAsn                              545550555560                                                                  LeuThrSerArgThrPheArgTyrThrAspPheSerAsnProPheSer                              565570575                                                                     PheArgAlaAsnProAspIleIleGlyIleSerGluGlnProLeuPhe                              580585590                                                                     GlyAlaGlySerIleSerSerGlyGluLeuTyrIleAspLysIleGlu                              595600605                                                                     IleIleLeuAlaAspAlaThrPheGluAlaGluSerAspLeuGluArg                              610615620                                                                     AlaGlnLysAlaValAsnAlaLeuPheThrSerSerAsnGlnIleGly                              625630635640                                                                  LeuLysThrAspValThrAspTyrHisIleAspGlnValSerAsnLeu                              645650655                                                                     ValAspCysLeuSerAspGluPheCysLeuAspGluLysArgGluLeu                              660665670                                                                     SerGluLysValLysHisAlaLysArgLeuSerAspGluArgAsnLeu                              675680685                                                                     LeuGlnAspProAsnPheArgGlyIleAsnArgGlnProAspArgGly                              690695700                                                                     TrpArgGlySerThrAspIleThrIleGlnGlyGlyAspAspValPhe                              705710715720                                                                  LysGluAsnTyrValThrLeuProGlyThrValAspGluCysTyrPro                              725730735                                                                     ThrTyrLeuTyrGlnLysIleAspGluSerLysLeuLysAlaTyrThr                              740745750                                                                     ArgTyrGluLeuArgGlyTyrIleGluAspSerGlnAspLeuGluIle                              755760765                                                                     TyrLeuIleArgTyrAsnAlaLysHisGluIleValAsnValProGly                              770775780                                                                     ThrGlySerLeuTrpProLeuSerAlaGlnSerProIleGlyLysCys                              785790795800                                                                  GlyGluProAsnArgCysAlaProHisLeuGluTrpAsnProAspLeu                              805810815                                                                     AspCysSerCysArgAspGlyGluLysCysAlaHisHisSerHisHis                              820825830                                                                     PheThrLeuAspIleAspValGlyCysThrAspLeuAsnGluAspLeu                              835840845                                                                     GlyValTrpValIlePheLysIleLysThrGlnAspGlyHisAlaArg                              850855860                                                                     LeuGlyAsnLeuGluPheLeuGluGluLysProLeuLeuGlyGluAla                              865870875880                                                                  LeuAlaArgValLysArgAlaGluLysLysTrpArgAspLysArgGlu                              885890895                                                                     LysLeuGlnLeuGluThrAsnIleValTyrLysGluAlaLysGluSer                              900905910                                                                     ValAspAlaLeuPheValAsnSerGlnTyrAspArgLeuGlnValAsp                              915920925                                                                     ThrAsnIleAlaMetIleHisAlaAlaAspLysArgValHisArgIle                              930935940                                                                     ArgGluAlaTyrLeuProGluLeuSerValIleProGlyValAsnAla                              945950955960                                                                  AlaIlePheGluGluLeuGluGlyArgIlePheThrAlaTyrSerLeu                              965970975                                                                     TyrAspAlaArgAsnValIleLysAsnGlyAspPheAsnAsnGlyLeu                              980985990                                                                     LeuCysTrpAsnValLysGlyHisValAspValGluGluGlnAsnAsn                              99510001005                                                                   HisArgSerValLeuValIleProGluTrpGluAlaGluValSerGln                              101010151020                                                                  GluValArgValCysProGlyArgGlyTyrIleLeuArgValThrAla                              1025103010351040                                                              TyrLysGluGlyTyrGlyGluGlyCysValThrIleHisGluIleGlu                              104510501055                                                                  AspAsnThrAspGluLeuLysPheSerAsnCysValGluGluGluVal                              106010651070                                                                  TyrProAsnAsnThrValThrCysAsnAsnTyrThrGlyThrGlnGlu                              107510801085                                                                  GluTyrGluGlyThrTyrThrSerArgAsnGlnGlyTyrAspGluAla                              109010951100                                                                  TyrGlyAsnAsnProSerValProAlaAspTyrAlaSerValTyrGlu                              1105111011151120                                                              GluLysSerTyrThrAspGlyArgArgGluAsnProCysGluSerAsn                              112511301135                                                                  ArgGlyTyrGlyAspTyrThrProLeuProAlaGlyTyrValThrLys                              114011451150                                                                  AspLeuGluTyrPheProGluThrAspLysValTrpIleGluIleGly                              115511601165                                                                  GluThrGluGlyThrPheIleValAspSerValGluLeuLeuLeuMet                              117011751180                                                                  GluGlu                                                                        1185                                                                          (2) INFORMATION FOR SEQ ID NO: 7:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 3579 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: unknown                                                         (ii) MOLECULE TYPE: cDNA                                                      (iii) HYPOTHETICAL: NO                                                        (iii) ANTI-SENSE: NO                                                          (vi) ORIGINAL SOURCE:                                                         (A) ORGANISM: Hybrid toxin                                                    (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..3579                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7:                                      ATGGATAACAATCCGAACATCAATGAATGCATTCCTTATAATTGTTTA48                            MetAspAsnAsnProAsnIleAsnGluCysIleProTyrAsnCysLeu                              151015                                                                        AGTAACCCTGAAGTAGAAGTATTAGGTGGAGAAAGAATAGAAACTGGT96                            SerAsnProGluValGluValLeuGlyGlyGluArgIleGluThrGly                              202530                                                                        TACACCCCAATCGATATTTCCTTGTCGCTAACGCAATTTCTTTTGAGT144                           TyrThrProIleAspIleSerLeuSerLeuThrGlnPheLeuLeuSer                              354045                                                                        GAATTTGTTCCCGGTGCTGGATTTGTGTTAGGACTAGTTGATATAATA192                           GluPheValProGlyAlaGlyPheValLeuGlyLeuValAspIleIle                              505560                                                                        TGGGGAATTTTTGGTCCCTCTCAATGGGACGCATTTCTTGTACAAATT240                           TrpGlyIlePheGlyProSerGlnTrpAspAlaPheLeuValGlnIle                              65707580                                                                      GAACAGTTAATTAACCAAAGAATAGAAGAATTCGCTAGGAACCAAGCC288                           GluGlnLeuIleAsnGlnArgIleGluGluPheAlaArgAsnGlnAla                              859095                                                                        ATTTCTAGATTAGAAGGACTAAGCAATCTTTATCAAATTTACGCAGAA336                           IleSerArgLeuGluGlyLeuSerAsnLeuTyrGlnIleTyrAlaGlu                              100105110                                                                     TCTTTTAGAGAGTGGGAAGCAGATCCTACTAATCCAGCATTAAGAGAA384                           SerPheArgGluTrpGluAlaAspProThrAsnProAlaLeuArgGlu                              115120125                                                                     GAGATGCGTATTCAATTCAATGACATGAACAGTGCCCTTACAACCGCT432                           GluMetArgIleGlnPheAsnAspMetAsnSerAlaLeuThrThrAla                              130135140                                                                     ATTCCTCTTTTTGCAGTTCAAAATTATCAAGTTCCTCTTTTATCAGTA480                           IleProLeuPheAlaValGlnAsnTyrGlnValProLeuLeuSerVal                              145150155160                                                                  TATGTTCAAGCTGCAAATTTACATTTATCAGTTTTGAGAGATGTTTCA528                           TyrValGlnAlaAlaAsnLeuHisLeuSerValLeuArgAspValSer                              165170175                                                                     GTGTTTGGACAAAGGTGGGGATTTGATGCCGCGACTATCAATAGTCGT576                           ValPheGlyGlnArgTrpGlyPheAspAlaAlaThrIleAsnSerArg                              180185190                                                                     TATAATGATTTAACTAGGCTTATTGGCAACTATACAGATCATGCTGTA624                           TyrAsnAspLeuThrArgLeuIleGlyAsnTyrThrAspHisAlaVal                              195200205                                                                     CGCTGGTACAATACGGGATTAGAGCGTGTATGGGGACCGGATTCTAGA672                           ArgTrpTyrAsnThrGlyLeuGluArgValTrpGlyProAspSerArg                              210215220                                                                     GATTGGATAAGATATAATCAATTTAGAAGAGAATTAACACTAACTGTA720                           AspTrpIleArgTyrAsnGlnPheArgArgGluLeuThrLeuThrVal                              225230235240                                                                  TTAGATATCGTTTCTCTATTTCCGAACTATGATAGTAGAACGTATCCA768                           LeuAspIleValSerLeuPheProAsnTyrAspSerArgThrTyrPro                              245250255                                                                     ATTCGAACAGTTTCCCAATTAACAAGAGAAATTTATACAAACCCAGTA816                           IleArgThrValSerGlnLeuThrArgGluIleTyrThrAsnProVal                              260265270                                                                     TTAGAAAATTTTGATGGTAGTTTTCGAGGCTCGGCTCAGGGCATAGAA864                           LeuGluAsnPheAspGlySerPheArgGlySerAlaGlnGlyIleGlu                              275280285                                                                     GGAAGTATTAGGAGTCCACATTTGATGGATATACTTAACAGTATAACC912                           GlySerIleArgSerProHisLeuMetAspIleLeuAsnSerIleThr                              290295300                                                                     ATCTATACGGATGCTCATAGAGGAGAATATTATTGGTCAGGGCATCAA960                           IleTyrThrAspAlaHisArgGlyGluTyrTyrTrpSerGlyHisGln                              305310315320                                                                  ATAATGGCTTCTCCTGTAGGGTTTTCGGGGCCAGAATTCACTTTTCCG1008                          IleMetAlaSerProValGlyPheSerGlyProGluPheThrPhePro                              325330335                                                                     CTATATGGAACTATGGGAAATGCAGCTCCACAACAACGTATTGTTGCT1056                          LeuTyrGlyThrMetGlyAsnAlaAlaProGlnGlnArgIleValAla                              340345350                                                                     CAACTAGGTCAGGGCGTGTATAGAACATTATCGTCCACTTTATATAGA1104                          GlnLeuGlyGlnGlyValTyrArgThrLeuSerSerThrLeuTyrArg                              355360365                                                                     AGACCTTTTAATATAGGGATAAATAATCAACAACTATCTGTTCTTGAC1152                          ArgProPheAsnIleGlyIleAsnAsnGlnGlnLeuSerValLeuAsp                              370375380                                                                     GGGACAGAATTTGCTTATGGAACCTCCTCAAATTTGCCATCCGCTGTA1200                          GlyThrGluPheAlaTyrGlyThrSerSerAsnLeuProSerAlaVal                              385390395400                                                                  TACAGAAAAAGCGGAACGGTAGATTCGCTGGATGAAATACCGCCACAG1248                          TyrArgLysSerGlyThrValAspSerLeuAspGluIleProProGln                              405410415                                                                     AATAACAACGTGCCACCTAGGCAAGGATTTAGTCATCGATTAAGCCAT1296                          AsnAsnAsnValProProArgGlnGlyPheSerHisArgLeuSerHis                              420425430                                                                     GTTTCAATGTTTCGTTCAGGCTTTAGTAATAGTAGTGTAAGTATAATA1344                          ValSerMetPheArgSerGlyPheSerAsnSerSerValSerIleIle                              435440445                                                                     AGAGCTCCTATGTTCTCTTGGATACATCGTAGTGCAACTCTTACAAAT1392                          ArgAlaProMetPheSerTrpIleHisArgSerAlaThrLeuThrAsn                              450455460                                                                     ACAATTGATCCAGAGAGAATTAATCAAATACCTTTAGTGAAAGGATTT1440                          ThrIleAspProGluArgIleAsnGlnIleProLeuValLysGlyPhe                              465470475480                                                                  AGAGTTTGGGGGGGCACCTCTGTCATTACAGGACCAGGATTTACAGGA1488                          ArgValTrpGlyGlyThrSerValIleThrGlyProGlyPheThrGly                              485490495                                                                     GGGGATATCCTTCGAAGAAATACCTTTGGTGATTTTGTATCTCTACAA1536                          GlyAspIleLeuArgArgAsnThrPheGlyAspPheValSerLeuGln                              500505510                                                                     GTCAATATTAATTCACCAATTACCCAAAGATACCGTTTAAGATTTCGT1584                          ValAsnIleAsnSerProIleThrGlnArgTyrArgLeuArgPheArg                              515520525                                                                     TACGCTTCCAGTAGGGATGCACGAGTTATAGTATTAACAGGAGCGGCA1632                          TyrAlaSerSerArgAspAlaArgValIleValLeuThrGlyAlaAla                              530535540                                                                     TCCACAGGAGTGGGAGGCCAAGTTAGTGTAAATATGCCTCTTCAGAAA1680                          SerThrGlyValGlyGlyGlnValSerValAsnMetProLeuGlnLys                              545550555560                                                                  ACTATGGAAATAGGGGAGAACTTAACATCTAGAACATTTAGATATACC1728                          ThrMetGluIleGlyGluAsnLeuThrSerArgThrPheArgTyrThr                              565570575                                                                     GATTTTAGTAATCCTTTTTCATTTAGAGCTAATCCAGATATAATTGGG1776                          AspPheSerAsnProPheSerPheArgAlaAsnProAspIleIleGly                              580585590                                                                     ATAAGTGAACAACCTCTATTTGGTGCAGGTTCTATTAGTAGCGGTGAA1824                          IleSerGluGlnProLeuPheGlyAlaGlySerIleSerSerGlyGlu                              595600605                                                                     CTTTATATAGATAAAATTGAAATTATTCTAGCAGATGCAACATTTGAA1872                          LeuTyrIleAspLysIleGluIleIleLeuAlaAspAlaThrPheGlu                              610615620                                                                     GCAGAATCTGATTTAGAAAGAGCACAAAAGGCGGTGAATGCCCTGTTT1920                          AlaGluSerAspLeuGluArgAlaGlnLysAlaValAsnAlaLeuPhe                              625630635640                                                                  ACTTCTTCCAATCAAATCGGGTTAAAAACCGATGTGACGGATTATCAT1968                          ThrSerSerAsnGlnIleGlyLeuLysThrAspValThrAspTyrHis                              645650655                                                                     ATTGATCAAGTATCCAATTTAGTGGATTGTTTATCAGATGAATTTTGT2016                          IleAspGlnValSerAsnLeuValAspCysLeuSerAspGluPheCys                              660665670                                                                     CTGGATGAAAAGCGAGAATTGTCCGAGAAAGTCAAACATGCGAAGCGA2064                          LeuAspGluLysArgGluLeuSerGluLysValLysHisAlaLysArg                              675680685                                                                     CTCAGTGATGAGCGGAATTTACTTCAAGATCCAAACTTCAGAGGGATC2112                          LeuSerAspGluArgAsnLeuLeuGlnAspProAsnPheArgGlyIle                              690695700                                                                     AATAGACAACCAGACCGTGGCTGGAGAGGAAGTACAGATATTACCATC2160                          AsnArgGlnProAspArgGlyTrpArgGlySerThrAspIleThrIle                              705710715720                                                                  CAAGGAGGAGATGACGTATTCAAAGAGAATTACGTCACACTACCGGGT2208                          GlnGlyGlyAspAspValPheLysGluAsnTyrValThrLeuProGly                              725730735                                                                     ACCGTTGATGAGTGCTATCCAACGTATTTATATCAGAAAATAGATGAG2256                          ThrValAspGluCysTyrProThrTyrLeuTyrGlnLysIleAspGlu                              740745750                                                                     TCGAAATTAAAAGCTTATACCCGTTATGAATTAAGAGGGTATATCGAA2304                          SerLysLeuLysAlaTyrThrArgTyrGluLeuArgGlyTyrIleGlu                              755760765                                                                     GATAGTCAAGACTTAGAAATCTATTTGATCCGTTACAATGCAAAACAC2352                          AspSerGlnAspLeuGluIleTyrLeuIleArgTyrAsnAlaLysHis                              770775780                                                                     GAAATAGTAAATGTGCCAGGCACGGGTTCCTTATGGCCGCTTTCAGCC2400                          GluIleValAsnValProGlyThrGlySerLeuTrpProLeuSerAla                              785790795800                                                                  CAAAGTCCAATCGGAAAGTGTGGAGAACCGAATCGATGCGCGCCACAC2448                          GlnSerProIleGlyLysCysGlyGluProAsnArgCysAlaProHis                              805810815                                                                     CTTGAATGGAATCCTGATCTAGATTGTTCCTGCAGAGACGGGGAAAAA2496                          LeuGluTrpAsnProAspLeuAspCysSerCysArgAspGlyGluLys                              820825830                                                                     TGTGCACATCATTCCCATCATTTCACCTTGGATATTGATGTTGGATGT2544                          CysAlaHisHisSerHisHisPheThrLeuAspIleAspValGlyCys                              835840845                                                                     ACAGACTTAAATGAGGACTTAGGTGTATGGGTGATATTCAAGATTAAG2592                          ThrAspLeuAsnGluAspLeuGlyValTrpValIlePheLysIleLys                              850855860                                                                     ACGCAAGATGGCCATGCAAGACTAGGGAATCTAGAGTTTCTCGAAGAG2640                          ThrGlnAspGlyHisAlaArgLeuGlyAsnLeuGluPheLeuGluGlu                              865870875880                                                                  AAACCATTATTAGGGGAAGCACTAGCTCGTGTGAAAAGAGCGGAGAAG2688                          LysProLeuLeuGlyGluAlaLeuAlaArgValLysArgAlaGluLys                              885890895                                                                     AAGTGGAGAGACAAACGAGAGAAACTGCAGTTGGAAACAAATATTGTT2736                          LysTrpArgAspLysArgGluLysLeuGlnLeuGluThrAsnIleVal                              900905910                                                                     TATAAAGAGGCAAAAGAATCTGTAGATGCTTTATTTGTAAACTCTCAA2784                          TyrLysGluAlaLysGluSerValAspAlaLeuPheValAsnSerGln                              915920925                                                                     TATGATAGATTACAAGTGGATACGAACATCGCGATGATTCATGCGGCA2832                          TyrAspArgLeuGlnValAspThrAsnIleAlaMetIleHisAlaAla                              930935940                                                                     GATAAACGCGTTCATAGAATCCGGGAAGCGTATCTGCCAGAGTTGTCT2880                          AspLysArgValHisArgIleArgGluAlaTyrLeuProGluLeuSer                              945950955960                                                                  GTGATTCCAGGTGTCAATGCGGCCATTTTCGAAGAATTAGAGGGACGT2928                          ValIleProGlyValAsnAlaAlaIlePheGluGluLeuGluGlyArg                              965970975                                                                     ATTTTTACAGCGTATTCCTTATATGATGCGAGAAATGTCATTAAAAAT2976                          IlePheThrAlaTyrSerLeuTyrAspAlaArgAsnValIleLysAsn                              980985990                                                                     GGCGATTTCAATAATGGCTTATTATGCTGGAACGTGAAAGGTCATGTA3024                          GlyAspPheAsnAsnGlyLeuLeuCysTrpAsnValLysGlyHisVal                              99510001005                                                                   GATGTAGAAGAGCAAAACAACCACCGTTCGGTCCTTGTTATCCCAGAA3072                          AspValGluGluGlnAsnAsnHisArgSerValLeuValIleProGlu                              101010151020                                                                  TGGGAGGCAGAAGTGTCACAAGAGGTTCGTGTCTGTCCAGGTCGTGGC3120                          TrpGluAlaGluValSerGlnGluValArgValCysProGlyArgGly                              1025103010351040                                                              TATATCCTTCGTGTCACAGCATATAAAGAGGGATATGGAGAGGGCTGC3168                          TyrIleLeuArgValThrAlaTyrLysGluGlyTyrGlyGluGlyCys                              104510501055                                                                  GTAACGATCCATGAGATCGAAGACAATACAGACGAACTGAAATTCAGC3216                          ValThrIleHisGluIleGluAspAsnThrAspGluLeuLysPheSer                              106010651070                                                                  AACTGTGTAGAAGAGGAAGTATATCCAAACAACACAGTAACGTGTAAT3264                          AsnCysValGluGluGluValTyrProAsnAsnThrValThrCysAsn                              107510801085                                                                  AATTATACTGGGACTCAAGAAGAATATGAGGGTACGTACACTTCTCGT3312                          AsnTyrThrGlyThrGlnGluGluTyrGluGlyThrTyrThrSerArg                              109010951100                                                                  AATCAAGGATATGACGAAGCCTATGGTAATAACCCTTCCGTACCAGCT3360                          AsnGlnGlyTyrAspGluAlaTyrGlyAsnAsnProSerValProAla                              1105111011151120                                                              GATTACGCTTCAGTCTATGAAGAAAAATCGTATACAGATGGACGAAGA3408                          AspTyrAlaSerValTyrGluGluLysSerTyrThrAspGlyArgArg                              112511301135                                                                  GAGAATCCTTGTGAATCTAACAGAGGCTATGGGGATTACACACCACTA3456                          GluAsnProCysGluSerAsnArgGlyTyrGlyAspTyrThrProLeu                              114011451150                                                                  CCGGCTGGTTATGTAACAAAGGATTTAGAGTACTTCCCAGAGACCGAT3504                          ProAlaGlyTyrValThrLysAspLeuGluTyrPheProGluThrAsp                              115511601165                                                                  AAGGTATGGATTGAGATCGGAGAAACAGAAGGAACATTCATCGTGGAT3552                          LysValTrpIleGluIleGlyGluThrGluGlyThrPheIleValAsp                              117011751180                                                                  AGCGTGGAATTACTCCTTATGGAGGAA3579                                               SerValGluLeuLeuLeuMetGluGlu                                                   11851190                                                                      (2) INFORMATION FOR SEQ ID NO: 8:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1193 amino acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8:                                      MetAspAsnAsnProAsnIleAsnGluCysIleProTyrAsnCysLeu                              151015                                                                        SerAsnProGluValGluValLeuGlyGlyGluArgIleGluThrGly                              202530                                                                        TyrThrProIleAspIleSerLeuSerLeuThrGlnPheLeuLeuSer                              354045                                                                        GluPheValProGlyAlaGlyPheValLeuGlyLeuValAspIleIle                              505560                                                                        TrpGlyIlePheGlyProSerGlnTrpAspAlaPheLeuValGlnIle                              65707580                                                                      GluGlnLeuIleAsnGlnArgIleGluGluPheAlaArgAsnGlnAla                              859095                                                                        IleSerArgLeuGluGlyLeuSerAsnLeuTyrGlnIleTyrAlaGlu                              100105110                                                                     SerPheArgGluTrpGluAlaAspProThrAsnProAlaLeuArgGlu                              115120125                                                                     GluMetArgIleGlnPheAsnAspMetAsnSerAlaLeuThrThrAla                              130135140                                                                     IleProLeuPheAlaValGlnAsnTyrGlnValProLeuLeuSerVal                              145150155160                                                                  TyrValGlnAlaAlaAsnLeuHisLeuSerValLeuArgAspValSer                              165170175                                                                     ValPheGlyGlnArgTrpGlyPheAspAlaAlaThrIleAsnSerArg                              180185190                                                                     TyrAsnAspLeuThrArgLeuIleGlyAsnTyrThrAspHisAlaVal                              195200205                                                                     ArgTrpTyrAsnThrGlyLeuGluArgValTrpGlyProAspSerArg                              210215220                                                                     AspTrpIleArgTyrAsnGlnPheArgArgGluLeuThrLeuThrVal                              225230235240                                                                  LeuAspIleValSerLeuPheProAsnTyrAspSerArgThrTyrPro                              245250255                                                                     IleArgThrValSerGlnLeuThrArgGluIleTyrThrAsnProVal                              260265270                                                                     LeuGluAsnPheAspGlySerPheArgGlySerAlaGlnGlyIleGlu                              275280285                                                                     GlySerIleArgSerProHisLeuMetAspIleLeuAsnSerIleThr                              290295300                                                                     IleTyrThrAspAlaHisArgGlyGluTyrTyrTrpSerGlyHisGln                              305310315320                                                                  IleMetAlaSerProValGlyPheSerGlyProGluPheThrPhePro                              325330335                                                                     LeuTyrGlyThrMetGlyAsnAlaAlaProGlnGlnArgIleValAla                              340345350                                                                     GlnLeuGlyGlnGlyValTyrArgThrLeuSerSerThrLeuTyrArg                              355360365                                                                     ArgProPheAsnIleGlyIleAsnAsnGlnGlnLeuSerValLeuAsp                              370375380                                                                     GlyThrGluPheAlaTyrGlyThrSerSerAsnLeuProSerAlaVal                              385390395400                                                                  TyrArgLysSerGlyThrValAspSerLeuAspGluIleProProGln                              405410415                                                                     AsnAsnAsnValProProArgGlnGlyPheSerHisArgLeuSerHis                              420425430                                                                     ValSerMetPheArgSerGlyPheSerAsnSerSerValSerIleIle                              435440445                                                                     ArgAlaProMetPheSerTrpIleHisArgSerAlaThrLeuThrAsn                              450455460                                                                     ThrIleAspProGluArgIleAsnGlnIleProLeuValLysGlyPhe                              465470475480                                                                  ArgValTrpGlyGlyThrSerValIleThrGlyProGlyPheThrGly                              485490495                                                                     GlyAspIleLeuArgArgAsnThrPheGlyAspPheValSerLeuGln                              500505510                                                                     ValAsnIleAsnSerProIleThrGlnArgTyrArgLeuArgPheArg                              515520525                                                                     TyrAlaSerSerArgAspAlaArgValIleValLeuThrGlyAlaAla                              530535540                                                                     SerThrGlyValGlyGlyGlnValSerValAsnMetProLeuGlnLys                              545550555560                                                                  ThrMetGluIleGlyGluAsnLeuThrSerArgThrPheArgTyrThr                              565570575                                                                     AspPheSerAsnProPheSerPheArgAlaAsnProAspIleIleGly                              580585590                                                                     IleSerGluGlnProLeuPheGlyAlaGlySerIleSerSerGlyGlu                              595600605                                                                     LeuTyrIleAspLysIleGluIleIleLeuAlaAspAlaThrPheGlu                              610615620                                                                     AlaGluSerAspLeuGluArgAlaGlnLysAlaValAsnAlaLeuPhe                              625630635640                                                                  ThrSerSerAsnGlnIleGlyLeuLysThrAspValThrAspTyrHis                              645650655                                                                     IleAspGlnValSerAsnLeuValAspCysLeuSerAspGluPheCys                              660665670                                                                     LeuAspGluLysArgGluLeuSerGluLysValLysHisAlaLysArg                              675680685                                                                     LeuSerAspGluArgAsnLeuLeuGlnAspProAsnPheArgGlyIle                              690695700                                                                     AsnArgGlnProAspArgGlyTrpArgGlySerThrAspIleThrIle                              705710715720                                                                  GlnGlyGlyAspAspValPheLysGluAsnTyrValThrLeuProGly                              725730735                                                                     ThrValAspGluCysTyrProThrTyrLeuTyrGlnLysIleAspGlu                              740745750                                                                     SerLysLeuLysAlaTyrThrArgTyrGluLeuArgGlyTyrIleGlu                              755760765                                                                     AspSerGlnAspLeuGluIleTyrLeuIleArgTyrAsnAlaLysHis                              770775780                                                                     GluIleValAsnValProGlyThrGlySerLeuTrpProLeuSerAla                              785790795800                                                                  GlnSerProIleGlyLysCysGlyGluProAsnArgCysAlaProHis                              805810815                                                                     LeuGluTrpAsnProAspLeuAspCysSerCysArgAspGlyGluLys                              820825830                                                                     CysAlaHisHisSerHisHisPheThrLeuAspIleAspValGlyCys                              835840845                                                                     ThrAspLeuAsnGluAspLeuGlyValTrpValIlePheLysIleLys                              850855860                                                                     ThrGlnAspGlyHisAlaArgLeuGlyAsnLeuGluPheLeuGluGlu                              865870875880                                                                  LysProLeuLeuGlyGluAlaLeuAlaArgValLysArgAlaGluLys                              885890895                                                                     LysTrpArgAspLysArgGluLysLeuGlnLeuGluThrAsnIleVal                              900905910                                                                     TyrLysGluAlaLysGluSerValAspAlaLeuPheValAsnSerGln                              915920925                                                                     TyrAspArgLeuGlnValAspThrAsnIleAlaMetIleHisAlaAla                              930935940                                                                     AspLysArgValHisArgIleArgGluAlaTyrLeuProGluLeuSer                              945950955960                                                                  ValIleProGlyValAsnAlaAlaIlePheGluGluLeuGluGlyArg                              965970975                                                                     IlePheThrAlaTyrSerLeuTyrAspAlaArgAsnValIleLysAsn                              980985990                                                                     GlyAspPheAsnAsnGlyLeuLeuCysTrpAsnValLysGlyHisVal                              99510001005                                                                   AspValGluGluGlnAsnAsnHisArgSerValLeuValIleProGlu                              101010151020                                                                  TrpGluAlaGluValSerGlnGluValArgValCysProGlyArgGly                              1025103010351040                                                              TyrIleLeuArgValThrAlaTyrLysGluGlyTyrGlyGluGlyCys                              104510501055                                                                  ValThrIleHisGluIleGluAspAsnThrAspGluLeuLysPheSer                              106010651070                                                                  AsnCysValGluGluGluValTyrProAsnAsnThrValThrCysAsn                              107510801085                                                                  AsnTyrThrGlyThrGlnGluGluTyrGluGlyThrTyrThrSerArg                              109010951100                                                                  AsnGlnGlyTyrAspGluAlaTyrGlyAsnAsnProSerValProAla                              1105111011151120                                                              AspTyrAlaSerValTyrGluGluLysSerTyrThrAspGlyArgArg                              112511301135                                                                  GluAsnProCysGluSerAsnArgGlyTyrGlyAspTyrThrProLeu                              114011451150                                                                  ProAlaGlyTyrValThrLysAspLeuGluTyrPheProGluThrAsp                              115511601165                                                                  LysValTrpIleGluIleGlyGluThrGluGlyThrPheIleValAsp                              117011751180                                                                  SerValGluLeuLeuLeuMetGluGlu                                                   11851190                                                                      __________________________________________________________________________

We claim:
 1. A purified fragment of a Bacillus thuringiensis toxincomprising amino acids 1-620 of SEQ ID NO:6, or a purified fragment of aBacillus thuringiensis toxin comprising amino acids 1-620 of SEQ ID NO:6wherein at least one of the following substitutions is present:Ile atposition 609 is replaced with Leu; Ala at position 618 is replaced withGlu; Ser at position 620 is replaced with Tyr.
 2. A purified fragment ofa Bacillus thuringiensis toxin comprising amino acids 1-627 of SEQ IDNO:8, or a purified fragment of a Bacillus thuringiensis toxincomprising amino acids 1-627 of SEQ ID NO:8 wherein at least one of thefollowing substitutions is present:Ile at position 617 is replaced withLeu; Ala at position 625 is replaced with Glu; Ser at position 627 isreplaced with Tyr.
 3. A purified fragment of a Bacillus thuringiensistoxin which has at least 95% sequence identity with, and has the sameinsecticidal specificity and substantially the same insecticidalactivity as the toxin fragment of claim 1 or
 2. 4. An insecticidalcomposition comprising the toxin fragment of claim 1 or
 2. 5. A processfor controlling insects comprising exposing them to the insecticidalcomposition of claim
 4. 6. A purified and isolated DNA encoding aprotein comprising the sequence of the toxin fragment of claim 1 or 2.7. A purified and isolated DNA comprising the sequence of nucleotides1-1860 of SEQ ID NO:5 or nucleotides 1-1881 of SEQ ID NO:7.
 8. Apurified and isolated DNA according to claim 6 or 7 which furtherencodes a protein having herbicide resistance, plant-growth promoting,anti-fungal antibacterial antiviral and/or anti-nematode properties. 9.A purified and isolated DNA according to claim 6 or 7 which is modifiedto optimize expression in a heterologous host, said modificationsselected from the group consisting of codon optimization for theintended host and removal of known mRNA instability motifs orpolyadenylation signals.
 10. A purified and isolated DNA according toclaim 8 which is modified to optimize expression in a heterologous host,said modifications selected from the group consisting of codonoptimization for the intended host and removal of known mRNA instabilitymotifs or polyadenylation signals.
 11. A recombinant vector comprising aDNA of claim 6, 7, 8, 9 or
 10. 12. An isolated cell transformed with aDNA of claim 6, 7, 8, 9 or
 10. 13. A method of producing a protein byexpressing the DNA of claim 6, 7, 8, 9 or
 10. 14. An isolated celltransformed with a vector of claim
 11. 15. An insecticidal compositioncomprising the isolated cell of claim
 12. 16. A process for controllinginsects comprising exposing them to the insecticidal composition ofclaim
 15. 17. Plants transformed with a DNA of claim 6, 7, 8, 9 or 10,the progeny of such plants which contain the DNA stably incorporated andheritable in a Mendelian manner.
 18. Seeds of the plants of claim 17.